Ig superfamily `dlair` receptors expressed in monocytes

ABSTRACT

DNA clones encoding a receptor in the Ig superfamily and a related soluble variant have been isolated from a human monocyte library. The invention provides receptor polypeptides, nucleic acids encoding them, expression vectors, and transformed cells for recombinant production of the polypeptides.

This filing is a conversion of, and claims benefit of priority to,provisional U.S. Patent Applications U.S. Ser. No. 60/032,252, filedDec. 6, 1996; U.S. Ser. No. 60/033,181, filed Dec. 16, 1996; and U.S.Ser. No. 60/041,279, filed Mar. 21, 1997, each of which is incorporatedherein by reference.

FIELD OF THE INVENTION

The present invention contemplates compositions related to genes foundin monocyte cells, cells which function in the immune system. Thesegenes function in controlling development, differentiation, and/orphysiology of the mammalian immune system. In particular, theapplication provides nucleic acids, proteins, antibodies, and methods ofusing them.

BACKGROUND OF THE INVENTION

The circulating component of the mammalian circulatory system comprisesvarious cell types, including red and white blood cells of the erythroidand myeloid cell lineages. See, e.g., Rapaport (1987) Introduction toHematology (2d ed.) Lippincott, Philadelphia, Pa.; Jandl (1987) Blood:Textbook of Hematology, Little, Brown and Co., Boston, Mass.; and Paul(ed.) (1993) Fundamental Immunology (3d ed.) Raven Press, N.Y.

Monocytes are phagocytic cells that belong to the mononuclear phagocytesystem and reside in the circulation. See Roitt (ed) Encyclopedia ofImmunology Academic Press, San Diego. These cells originate in the bonemarrow and remain only a short time in the marrow compartment once theydifferentiate. They then enter the circulation and can remain there fora relatively long period of time, e.g., a few days. The monocytes canenter the tissues and body cavities by the process designateddiapedesis, where they differentiate into macrophages and possibly intodendritic cells. In an inflammatory response, the number of monocytes inthe circulation may double, and many of the increased number ofmonocytes diapedese to the site of inflammation.

Antigen presentation refers to the cellular events in which aproteinaceous antigen is taken up, processed by antigen presenting cells(APC), and then recognized to initiate an immune response. The mostactive antigen presenting cells have been characterized as themacrophages, which are direct developmental products from monocytes;dendritic cells; and certain B cells.

Macrophages are found in most tissues and are highly active ininternalization of a wide variety of protein antigens andmicroorganisms. They have a highly developed endocytic activity, andsecrete many products important in the initiation of an immune response.For this reason, it is believed that many genes expressed by monocytesor induced by monocyte activation are likely to be important in antigenuptake, processing, presentation, or regulation of the resulting immuneresponse.

However, monocytes are poorly characterized, both in terms of proteinsthey express, and many of their functions and mechanisms of action,including their activated states. In particular, the processes andmechanisms related to the initiation of an immune response, includingantigen processing and presentation, remain unclear. The absence ofknowledge about the structural, biological, and physiological propertiesof these cells limits their understanding. Thus, medical conditionswhere regulation, development, or physiology of antigen presenting cellsis unusual remain unmanageable.

SUMMARY OF THE INVENTION

The present invention is based, in part, upon the discovery of variousgenes isolated from activated monocytes. These molecules have beendesignated FDF03 (a type I transmembrane protein with Ig-likeextracellular portion); YE01 (an Fc gamma/alpha-like receptor); andKTE03 class (cell surface receptors exhibiting Ig-like domains),represented by YYB01, YYB04 related, KLTM63, KT66, and KLM67embodiments.

The invention provides various compositions of matter selected from: asubstantially pure or recombinant FDF03 protein or peptide exhibiting atleast about 85% sequence identity over a length of at least about 12amino acids to mature SEQ ID NO: 2 or 4; a natural sequence FDF03 of SEQID NO: 2 or 4; a fusion protein comprising FDF03 sequence; asubstantially pure or recombinant YE01 protein or peptide exhibiting atleast about 85% sequence identity over a length of at least about 12amino acids to mature SEQ ID NO: 6, 8, or 10; a natural sequence YE01 ofSEQ ID NO: 6, 8, or 10; a fusion protein comprising YE01 sequence; asubstantially pure or recombinant KTE03 protein or peptide exhibiting atleast about 85% sequence identity over a length of at least about 12amino acids to SEQ ID NO: 12, 14, 16, 18, 20, or 22; a natural sequenceKTE03 of SEQ ID NO: 12, 14, 16, 18, 20, or 22; or a fusion proteincomprising KTE03 sequence. Preferably, the substantially pure orisolated protein comprises a segment exhibiting sequence identity to acorresponding portion of a FDF03, YE01, or KTE03, wherein: the homologyis at least about 90% identity and the portion is at least about 9 aminoacids; the homology is at least about 80% identity and the portion is atleast about 17 amino acids; or the homology is at least about 70%identity and the portion is at least about 25 amino acids. In otherforms, the invention provides such composition of matter, wherein the:FDF03 comprises a mature sequence of Table 1; YE01 comprises a maturesequence of Table 2; KTE03 comprises a mature sequence of Table 3; orthe protein or peptide: is from a warm blooded animal selected from amammal, including a primate or rodent; comprises at least onepolypeptide segment of SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, or22; exhibits a plurality of portions exhibiting the identity; is anatural allelic variant of FDF03, YE01, or KTE03; has a length at leastabout 30 amino acids; exhibits at least two non-overlapping epitopeswhich are specific for a mammalian FDF03, YE01, or KTE03; exhibits asequence identity at least about 90% over a length of at least about 20amino acids to a rodent FDF03, YE01, or KTE03; exhibits at least twonon-overlapping epitopes which are specific for a primate FDF03, YE01,or KTE03; exhibits a sequence identity at least about 90% over a lengthof at least about 20 amino acids to a primate FDF03, YE01, or KTE03; isglycosylated; has a molecular weight of at least 7 kD with naturalglycosylation; is a synthetic polypeptide; is attached to a solidsubstrate; is conjugated to another chemical moiety; is a 5-fold or lesssubstitution from natural sequence; or is a deletion or insertionvariant from a natural sequence.

Other compositions include those comprising: a sterile FDF03 protein orpeptide; the FDF03 protein or peptide and a carrier, wherein the carrieris: an aqueous compound, including water, saline, and/or buffer; and/orformulated for oral, rectal, nasal, topical, or parenteraladministration; a sterile YE01 protein or peptide; the YE01 protein orpeptide and a carrier, wherein the carrier is: an aqueous compound,including water, saline, and/or buffer; and/or formulated for oral,rectal, nasal, topical, or parenteral administration; a sterile KTE03protein or peptide; or the KTE03 protein or peptide and a carrier,wherein the carrier is: an aqueous compound, including water, saline,and/or buffer; and/or formulated for oral, rectal, nasal, topical, orparenteral administration.

In fusion protein embodiments, the invention provides those whichcomprise: mature protein sequence of Table 1, 2, or 3; a detection orpurification tag, including a FLAG, His6, or Ig sequence; or sequence ofanother cell surface protein.

Various kits include those comprising a protein or polypeptide, and: acompartment comprising the protein or polypeptide; and/or instructionsfor use or disposal of reagents in the kit.

Antibodies and binding compounds include those comprising an antigenbinding portion from an antibody, which specifically binds to a naturalFDF03, YE01, or KTE03 protein, wherein: the protein is a primateprotein; the binding compound is an Fv, Fab, or Fab2 fragment; thebinding compound is conjugated to another chemical moiety; or theantibody: is raised against a peptide sequence of a mature polypeptideof Table 1, 2, or 3; is raised against a mature FDF03, YE01, or KTE03;is raised to a purified FDF03, YE01, or KTE03; is immunoselected; is apolyclonal antibody; binds to a denatured FDF03, YE01, or KTE03;exhibits a Kd to antigen of at least 30 μM; is attached to a solidsubstrate, including a bead or plastic membrane; is in a sterilecomposition; or is detectably labeled, including a radioactive orfluorescent label. A kit comprising the binding compound is providedincluding, e.g., the binding compound and: a compartment comprising thebinding compound; and/or instructions for use or disposal of reagents inthe kit. Preferably, the kit is capable of making a qualitative orquantitative analysis.

Various other compositions include those comprising: a sterile bindingcompound; or the binding compound and a carrier, wherein the carrier is:an aqueous compound, including water, saline, and/or buffer; and/orformulated for oral, rectal, nasal, topical, or parenteraladministration.

Nucleic acid embodiments include an isolated or recombinant nucleic acidencoding a protein or peptide or fusion protein as described, wherein:the protein is from a mammal, including a primate; or the nucleic acid:encodes an antigenic peptide sequence of Table 1, 2, or 3; encodes aplurality of antigenic peptide sequences of Table 1, 2, or 3; exhibitsat least about 80% identity to a natural cDNA encoding the segment; isan expression vector; further comprises an origin of replication; isfrom a natural source; comprises a detectable label; comprises syntheticnucleotide sequence; is less than 6 kb, preferably less than 3 kb; isfrom a mammal, including a primate; comprises a natural full lengthcoding sequence; is a hybridization probe for a gene encoding theprotein; or is a PCR primer, PCR product, or mutagenesis primer.

Various cells are provided, including those comprising a describedrecombinant nucleic acid. Preferably, the cell is: a prokaryotic cell; aeukaryotic cell; a bacterial cell; a yeast cell; an insect cell; amammalian cell; a mouse cell; a primate cell; or a human cell. Kits withsuch nucleic acids include those with the nucleic acid and: acompartment comprising the nucleic acid; a compartment furthercomprising a FDF03, YE01, or KTE03 protein or polypeptide; and/orinstructions for use or disposal of reagents in the kit. Preferably, thekit is capable of making a qualitative or quantitative analysis.

Other nucleic acids include those which: hybridize under wash conditionsof 30° C. and less than 2M salt to the coding portions of SEQ ID NO: 1or 3; hybridize under wash conditions of 30° C. and less than 2 M saltto the coding portions of SEQ ID NO: 5, 7, or 9; hybridize under washconditions of 30° C. and less than 2M salt to the coding portions of SEQID NO: 11, 13, 15, 17, 19, or 21; exhibit at least about 85% identityover a stretch of at least about 30 nucleotides to a primate FDF03;exhibit at least about 85% identity over a stretch of at least about 30nucleotides to a primate YE01; or exhibit at least about 85% identityover a stretch of at least about 30 nucleotides to a primate KTE03. Inpreferred embodiments, the wash conditions are at 45° C. and/or 500 mMsalt; or at 55° C. and/or 150 mM salt; or the identity is at least 90%and/or the stretch is at least 55 nucleotides; or the identity is atleast 95% and/or the stretch is at least 75 nucleotides.

The invention further provides a method of modulating physiology ordevelopment of a cell or tissue culture cell comprising contacting thecell with an agonist or antagonist of a FDF03, YE01, or KTE03. Inpreferred embodiments, the cell is a leukocyte, and the antagonist is toYE01 and is a monoclonal antibody which binds to DLAIR-1.

DETAILED DESCRIPTION

    ______________________________________                                         OUTLINE                                                                  

    ______________________________________                                         I.         General                                                             II.  Definitions                                                              III. Nucleic Acids                                                            IV.  Making Proteins                                                          V.   Antibodies                                                               VI.  Purified Proteins                                                        VII. Physical Variants                                                        VIII. Binding Agent:Monocyte Protein Complexes                                IX.  Uses                                                                     X.   Kits                                                                     XI.  Binding Partner Isolation                                              ______________________________________                                    

I. General

The present invention provides DNA sequences encoding mammalian proteinsexpressed on monocytes. For a review of monocytes and their functions,see, e.g., Gallin, et al. (eds. 1988) Inflammation: Basic Principles andClinical Correlates Raven Press, NY; van Furth (ed. 1985) MononuclearPhagocytes: Characteristics, Physiology and Function Martinus Nijhoff,Dordrecht, Netherlands.

Specific human embodiments of these proteins are provided below. Thedescriptions below are directed, for exemplary purposes, to humanmonocyte genes, but are likewise applicable to structurally, e.g.,sequence, related embodiments from other sources or mammalian species,including polymorphic or individual variants. These will include, e.g.,proteins which exhibit a relatively few changes in sequence, e.g., lessthan about 5%, and number, e.g., less than 20 residue substitutions,typically less than 15, preferably less than 10, and more preferablyless than 5 substitutions. These will also include versions which aretruncated from full length, as described, and fusion proteins containingsubstantial segments of these sequences.

II. Definitions

The term "binding composition" refers to molecules that bind withspecificity to a these monocyte proteins, e.g., in an antibody-antigeninteraction, or compounds, e.g., proteins, which specifically associatewith the respective protein. Typically, the association will be in anatural physiologically relevant protein-protein interaction, eithercovalent or non-covalent, and may include members of a multiproteincomplex, including carrier compounds or dimerization partners. Themolecule may be a polymer, or chemical reagent. A functional analog maybe a protein with structural modifications, or may be a wholly unrelatedmolecule, e.g., which has a molecular shape which interacts with theappropriate interacting determinants. The variants may serve as agonistsor antagonists of the protein, see, e.g., Goodman, et al. (eds.) (1990)Goodman & Gilman's: The Pharmacological Bases of Therapeutics (8th ed.)Pergamon Press, Tarrytown, N.Y.

The term "binding agent:monocyte protein complex", as used herein,refers to a complex of a binding agent and the monocyte protein.Specific binding of the binding agent means that the binding agent has aspecific binding site that recognizes a site on the respective monocyteprotein. For example, antibodies raised to the monocyte protein andrecognizing an epitope on the monocyte protein are capable of forming abinding agent:monocyte protein complex by specific binding. Typically,the formation of a binding agent:monocyte protein complex allows themeasurement of monocyte protein in a mixture of other proteins andbiologics. The term "antibody:monocyte protein complex" refers to abinding agent:monocyte protein complex in which the binding agent is anantibody. The antibody may be monoclonal, polyclonal or even an antigenbinding fragment of an antibody.

"Homologous" nucleic acid sequences, when compared, exhibit significantsimilarity. The standards for homology in nucleic acids are eithermeasures for homology generally used in the art by sequence comparisonand/or phylogenetic relationship, or based upon hybridizationconditions. Hybridization conditions are described in greater detailbelow.

An "isolated" nucleic acid is a nucleic acid, e.g., an RNA, DNA, or amixed polymer, which is substantially separated from other componentswhich naturally accompany a native sequence, e.g., proteins and flankinggenomic sequences from the originating species. The term embraces anucleic acid sequence which has been removed from its naturallyoccurring environment, and includes recombinant or cloned DNA isolatesand chemically synthesized analogs or analogs biologically synthesizedby heterologous systems. A substantially pure molecule includes isolatedforms of the molecule. An isolated nucleic acid will generally be ahomogeneous composition of molecules, but will, in some embodiments,contain minor heterogeneity. This heterogeneity is typically found atthe polymer ends or portions not critical to a desired biologicalfunction or activity.

As used herein, the term "monocyte protein" shall encompass, when usedin a protein context, a protein having amino acid sequences as shown inSEQ ID NO: 2 or 4; 6, 8, or 10; or 12, 14, 16, 18, 20, or 22, or asignificant fragment of such a protein. It refers to a polypeptide whichinteracts with the respective monocyte protein specific bindingcomponents. These binding components, e.g., antibodies, typically bindto the monocyte protein with high affinity, e.g., at least about 100 nM,usually better than about 30 nM, preferably better than about 10 nM, andmore preferably at better than about 3 nM.

The term "polypeptide" or "protein" as used herein includes asignificant fragment or segment of said monocyte protein, andencompasses a stretch of amino acid residues of at least about 8 aminoacids, generally at least 10 amino acids, more generally at least 12amino acids, often at least 14 amino acids, more often at least 16 aminoacids, typically at least 18 amino acids, more typically at least 20amino acids, usually at least 22 amino acids, more usually at least 24amino acids, preferably at least 26 amino acids, more preferably atleast 28 amino acids, and, in particularly preferred embodiments, atleast about 30 or more amino acids. Fragment or size limitationsapplicable for comparison to one group, e.g., to the FDF03, do notnecessarily imply similar size limitations on fragments for the others.

A "recombinant" nucleic acid is defined either by its method ofproduction or its structure. In reference to its method of production,e.g., a product made by a process, the process is use of recombinantnucleic acid techniques, e.g., involving human intervention in thenucleotide sequence, typically selection or production. Alternatively,it can be a nucleic acid made by generating a sequence comprising fusionof two fragments which are not naturally contiguous to each other, butis meant to exclude products of nature, e.g., naturally occurringmutants. Thus, for example, products made by transforming cells with anynon-naturally occurring vector is encompassed, as are nucleic acidscomprising sequence derived using any synthetic oligonucleotide process.Such is often done to replace a codon with a redundant codon encodingthe same or a conservative amino acid, while typically introducing orremoving a sequence recognition site. Alternatively, it is performed tojoin together nucleic acid segments of desired functions to generate asingle genetic entity comprising a desired combination of functions notfound in the commonly available natural forms. Restriction enzymerecognition sites are often the target of such artificial manipulations,but other site specific targets, e.g., promoters, DNA replication sites,regulation sequences, control sequences, or other useful features may beincorporated by design. A similar concept is intended for a recombinant,e.g., fusion, polypeptide. Specifically included are synthetic nucleicacids which, by genetic code redundancy, encode polypeptides similar tofragments of these antigens, and fusions of sequences from variousdifferent species variants.

"Solubility" is reflected by sedimentation measured in Svedberg units,which are a measure of the sedimentation velocity of a molecule underparticular conditions. The determination of the sedimentation velocitywas classically performed in an analytical ultracentrifuge, but istypically now performed in a standard ultracentrifuge. See, Freifelder(1982) Physical Biochemistry (2d ed.) W. H. Freeman & Co., SanFrancisco, Calif.; and Cantor and Schimmel (1980) Biophysical Chemistryparts 1-3, W. H. Freeman & Co., San Francisco, Calif. As a crudedetermination, a sample containing a putatively soluble polypeptide isspun in a standard full sized ultracentrifuge at about 50K rpm for about10 minutes, and soluble molecules will remain in the supernatant. Asoluble particle or polypeptide will typically be less than about 30S,more typically less than about 15S, usually less than about 10S, moreusually less than about 6S, and, in particular embodiments, preferablyless than about 4S, and more preferably less than about 3S. Solubilityof a polypeptide or fragment depends upon the environment and thepolypeptide. Many parameters affect polypeptide solubility, includingtemperature, electrolyte environment, size and molecular characteristicsof the polypeptide, and nature of the solvent. Typically, thetemperature at which the polypeptide is used ranges from about 4° C. toabout 65° C. Usually the temperature at use is greater than about 18° C.and more usually greater than about 22° C. For diagnostic purposes, thetemperature will usually be about room temperature or warmer, but lessthan the denaturation temperature of components in the assay. Fortherapeutic purposes, the temperature will usually be body temperature,typically about 37° C. for humans, though under certain situations thetemperature may be raised or lowered in situ or in vitro.

The size and structure of the polypeptide should generally be in asubstantially stable state, and usually not in a denatured state. Thepolypeptide may be associated with other polypeptides in a quaternarystructure, e.g., to confer solubility, or associated with lipids ordetergents in a manner which approximates natural lipid bilayerinteractions.

The solvent will usually be a biologically compatible buffer, of a typeused for preservation of biological activities, and will usuallyapproximate a physiological solvent. Usually the solvent will have aneutral pH, typically between about 5 and 10, and preferably about 7.5.On some occasions, a detergent will be added, typically a mildnon-denaturing one, e.g., CHS or CHAPS, or a low enough concentration asto avoid significant disruption of structural or physiologicalproperties of the protein.

"Substantially pure" typically means that the protein is isolated fromother contaminating proteins, nucleic acids, and other biologicalsderived from the original source organism. Purity, or "isolation" may beassayed by standard methods, and will ordinarily be at least about 50%pure, more ordinarily at least about 60% pure, generally at least about70% pure, more generally at least about 80% pure, often at least about85% pure, more often at least about 90% pure, preferably at least about95% pure, more preferably at least about 98% pure, and in most preferredembodiments, at least 99% pure.

"Substantial similarity" in the nucleic acid sequence comparison contextmeans either that the segments, or their complementary strands, whencompared, are identical when optimally aligned, with appropriatenucleotide insertions or deletions, in at least about 50% of thenucleotides, generally at least 56%, more generally at least 59%,ordinarily at least 62%, more ordinarily at least 65%, often at least68%, more often at least 71%, typically at least 74%, more typically atleast 77%, usually at least 80%, more usually at least about 85%,preferably at least about 90%, more preferably at least about 95 to 98%or more, and in particular embodiments, as high at about 99% or more ofthe nucleotides. Alternatively, substantial similarity exists when thesegments will hybridize under selective hybridization conditions, to astrand, or its complement, typically using a sequence derived from SEQID NO: 1 or 3; 5, 7, or 9; or 11, 13, 15, 17, 19, or 21. Typically,selective hybridization will occur when there is at least about 55%similarity over a stretch of at least about 30 nucleotides, preferablyat least about 65% over a stretch of at least about 25 nucleotides, morepreferably at least about 75%, and most preferably at least about 90%over about 20 nucleotides. See, e.g., Kanehisa (1984) Nucl. Acids Res.12:203-213. The length of similarity comparison, as described, may beover longer stretches, and in certain embodiments will be over a stretchof at least about 17 nucleotides, usually at least about 20 nucleotides,more usually at least about 24 nucleotides, typically at least about 28nucleotides, more typically at least about 40 nucleotides, preferably atleast about 50 nucleotides, and more preferably at least about 75 to 100or more nucleotides.

"Stringent conditions", in referring to homology or substantialsimilarity in the hybridization context, will be stringent combinedconditions of salt, temperature, organic solvents, and other parameters,typically those controlled in hybridization reactions. The combinationof parameters is more important than the measure of any singleparameter. See, e.g., Wetmur and Davidson (1968) by J. Mol. Biol.31:349-370. A nucleic acid probe which binds to a target nucleic acidunder stringent conditions is specific for said target nucleic acid.Such a probe is typically more than 11 nucleotides in length, and issufficiently identical or complementary to a target nucleic acid overthe region specified by the sequence of the probe to bind the targetunder stringent hybridization conditions.

Counterpart monocyte proteins from other mammalian species can be clonedand isolated by cross-species hybridization of closely related species.See, e.g., below. Similarity may be relatively low between distantlyrelated species, and thus hybridization of relatively closely relatedspecies is advisable. Alternatively, preparation of an antibodypreparation which exhibits less species specificity may be useful inexpression cloning approaches.

The phrase "specifically binds to an antibody" or "specificallyimmunoreactive with", when referring to a protein or peptide, refers toa binding reaction which is determinative of the presence of the proteinin the presence of a heterogeneous population of proteins and otherbiological components. Thus, under designated immunoassay conditions,the specified antibodies bind to a particular protein and do notsignificantly bind other proteins present in the sample. Specificbinding to an antibody under such conditions may require an antibodythat is selected for its specificity for a particular protein. Forexample, antibodies raised to the human monocyte protein immunogen withthe amino acid sequence depicted in SEQ ID NO: 2 can be selected toobtain antibodies specifically immunoreactive with that monocyte proteinand not with other proteins. These antibodies recognize proteins highlysimilar to the homologous human monocyte protein.

III. Nucleic Acids

These monocyte genes are specifically expressed on dendritic cells. Thepreferred embodiments, as disclosed, will be useful in standardprocedures to isolate genes from other species, e.g., warm bloodedanimals, such as birds and mammals. Cross hybridization will allowisolation of related proteins from individuals, strains, or species. Anumber of different approaches are available successfully to isolate asuitable nucleic acid clone based upon the information provided herein.Southern blot hybridization studies should identify homologous genes inother species under appropriate hybridization conditions.

Purified protein or defined peptides are useful for generatingantibodies by standard methods, as described below. Synthetic peptidesor purified protein can be presented to an immune system to generatepolyclonal and monoclonal antibodies. See, e.g., Coligan (1991) CurrentProtocols in Immunology Wiley/Greene, NY; and Harlow and Lane (1989)Antibodies: A Laboratory Manual Cold Spring Harbor Press, NY, which areincorporated herein by reference. Alternatively, a CD protein bindingcomposition can be useful as a specific binding reagent, and advantagecan be taken of its specificity of binding, for, e.g., purification of amonocyte protein.

The specific binding composition can be used for screening an expressionlibrary made from a cell line which expresses the respective monocyteprotein. Many methods for screening are available, e.g., standardstaining of surface expressed ligand, or by panning. Screening ofintracellular expression can also be performed by various staining orimmunofluorescence procedures. The binding compositions could be used toaffinity purify or sort out cells expressing the ligand.

                                      TABLE 1                                     __________________________________________________________________________    Sequence encoding a human FDF03 protein, containing Ig domains.                 The putative coding region runs from about 154 to 1062. See                   SEQ ID NO: 1 and 2. This 1249 bp clone was isolated from a                    monocyte cell library. A putative (hydrophobic) signal sequence              runs from -19 to about -1; a putative transmembrane (hydrophobic)             segment runs from about 178 to 199. The extracellular region is               probably about 170 amino acids, with a potential Ig-like domain               structure; the intracellular region is about 80 residues.                     Sequence analysis indicates similarity to GenBank clones H26010               and R50327 from humans.                                                      GTTTGGGGAA GGCTCCTGGC CCCCACAGCC CTCTTCGGAG CCTGAGCCCG GCTCTCCTCA                                                     60                                       - CTCACCTCAA CCCCCAGGCG GCCCCTCCAC AGGGCCCCTC TCCTGCCTGG ACGGCTCTGC                                                120                                      - TGGTCTCCCC GTCCCCTGGA GAAGAACAAG GCC ATG GGT CGG CCC CTG CTG CTG 174                                            Met Gly Arg Pro Leu Leu Leu                                                        -19             -15                  - CCC CTA CTG CCC CTG CTG CTG CCG CCA GCA TTT CTG CAG CCT AGT GGC 222                                               Pro Leu Leu Pro Leu Leu Leu Pro                                              Pro Ala Phe Leu Gln Pro Ser Gly                                                        -10                  -5                                                               1                       - TCC ACA GGA TCT GGT CCA AGC TAC CTT TAT GGG GTC ACT CAA CCA AAA 270                                               Ser Thr Gly Ser Gly Pro Ser Tyr                                              Leu Tyr Gly Val Thr Gln Pro Lys                                                  5                  10                                                              15                  20                                                  - CAC CTC TCA GCC TCC ATG GGT                                               GGC TCT GTG GAA ATC CCC TTC TCC                                               TTC 318                                 His Leu Ser Ala Ser Met Gly Gly Ser Val Glu Ile Pro Phe Ser Phe                                                                       25                                                              30                  35                                                      - TAT TAC CCC TGG GAG TTA GCC                                               ACA GCT CCC GAC GTG AGA ATA TCC                                               TGG 366                                 Tyr Tyr Pro Trp Glu Leu Ala Thr Ala Pro Asp Val Arg Ile Ser Trp                                                                   40                                                              45                  50                   - AGA CGG GGC CAC TTC CAC GGG CAG TCC TTC TAC AGC ACA AGG CCG CCT 414                                               Arg Arg Gly His Phe His Gly Gln                                              Ser Phe Tyr Ser Thr Arg Pro Pro                                                         55                  60                                                              65                       - TCC ATT CAC AAG GAT TAT GTG AAC CGG CTC TTT CTG AAC TGG ACA GAG 462                                               Ser Ile His Lys Asp Tyr Val Asn                                              Arg Leu Phe Leu Asn Trp Thr Glu                                                     70                  75                                                              80                           - GGT CAG AAG AGC GGC TTC CTC AGG ATC TCC AAC CTG CAG AAG CAG GAC 510                                               Gly Gln Lys Ser Gly Phe Leu Arg                                              Ile Ser Asn Leu Gln Lys Gln Asp                                                 85                  90                                                              95                 100                                                  - CAG TCT GTG TAT TTC TGC CGA                                               GTT GAG CTG GAC ACA CGG AGC TCA                                               GGG 558                                 Gln Ser Val Tyr Phe Cys Arg Val Glu Leu Asp Thr Arg Ser Ser Gly                                                                      105                                                             110                 115                                                      - AGG CAG CAG TGG CAG TCC ATC                                               GAG GGG ACC AAA CTC TCC ATC ACC                                               CAG 606                                 Arg Gln Gln Trp Gln Ser Ile Glu Gly Thr Lys Leu Ser Ile Thr Gln                                                                  120                                                              125                 130                  - GCT GTC ACG ACC ACC ACC CAG AGG CCC AGC AGC ATG ACT ACC ACC TGG 654                                               Ala Val Thr Thr Thr Thr Gln Arg                                              Pro Ser Ser Met Thr Thr Thr Trp                                                        135                 140                                                             145                       - AGG CTC AGT AGC ACA ACC ACC ACA ACC GGC CTC AGG GTC ACA CAG GGC 702                                               Arg Leu Ser Ser Thr Thr Thr Thr                                              Thr Gly Leu Arg Val Thr Gln Gly                                                    150                 155                                                             160                           - AAA CGA CGC TCA GAC TCT TGG CAC ATA AGT CTG GAG ACT GCT GTG GGG 750                                               Lys Arg Arg Ser Asp Ser Trp His                                              Ile Ser Leu Glu Thr Ala Val Gly                                                165                 170                                                             175                 180                                                  - GTG GCA GTG GCT GTC ACT GTG                                               CTC GGA ATC ATG ATT TTG GGA CTG                                               ATC 798                                 Val Ala Val Ala Val Thr Val Leu Gly Ile Met Ile Leu Gly Leu Ile                                                                      185                                                             190                 195                                                      - TGC CTC CTC AGG TGG AGG AGA                                               AGG AAA GGT CAG CAG CGG ACT AAA                                               GCC 846                                 Cys Leu Leu Arg Trp Arg Arg Arg Lys Gly Gln Gln Arg Thr Lys Ala                                                                  200                                                              205                 210                  - ACA ACC CCA GCC AGG GAA CCC TTC CAA AAC ACA GAG GAG CCA TAT GAG 894                                               Thr Thr Pro Ala Arg Glu Pro Phe                                              Gln Asn Thr Glu Glu Pro Tyr Glu                                                        215                 220                                                             225                       - AAT ATC AGG AAT GAA GGA CAA AAT ACA GAT CCC AAG CTA AAT CCC AAG 942                                               Asn Ile Arg Asn Glu Gly Gln Asn                                              Thr Asp Pro Lys Leu Asn Pro Lys                                                    230                 235                                                             240                           - GAT GAC GGC ATC GTA TAT GCT TCC CTT GCC CTC TCC AGC TCC ACC TCA 990                                               Asp Asp Gly Ile Val Tyr Ala Ser                                              Leu Ala Leu Ser Ser Ser Thr Ser                                                245                 250                                                             255                 260                                                  - CCC AGA GCA CCT CCC AGC CAC                                               CGT CCC CTC AAG AGC CCC CAG AAC                                               GAG 1038                                Pro Arg Ala Pro Pro Ser His Arg Pro Leu Lys Ser Pro Gln Asn Glu                                                                      265                                                             270                 275                                                      - ACC CTG TAC TCT GTC TTA AAG                                               GCC TAACCAATGG ACAGCCCTCT                                                     CAAGACTGAA 1092                         Thr Leu Tyr Ser Val Leu Lys Ala                                                           280                                                                - TGGTGAGGCC AGGTACAGTG GCGCACACCT GTAATCCCAG CTACTCTGAA GCCTGAGGCA                                                1152                                     - GAATCAAGTG AGCCCAGGAG TTCAGGGCCA GCTTTGATAA TGGAGCGAGA TGCCATCTCT                                                1212                                     - AGTTAAAAAT ATATATTAAC AATAAAGTAA CAAATTT 1249                               - A mouse counterpart partial sequence is (SEQ ID NO: 3 and 4):            CCCCAGTGTC CCTAGACAGA GCATCCTTGC CTTCCTGATG GCTTTGCTGA TCTCGCTTCC                                                     60                                       - CTGGAGGGAC TCCAGCC ATG GCT CAG GTC CTG CTT CTG CTC TCA TCA GGC 110                                                                   Met Ala Gln                                               Val Leu Leu Leu Leu Ser Ser Gly                                                                     1                                                           5                  10                                                       - TGT CTG CAT GCT GGA AAT TCA                                               GAA AGA TAC AAC AGA AAA AAT GGC                                               TTT 158                                 Cys Leu His Ala Gly Asn Ser Glu Arg Tyr Asn Arg Lys Asn Gly Phe                                                                   15                                                              20                  25                   - GGG GTC AAC CAA CCT GAA CGC TGC TCT GGA GTC CAG GGT GGC TCC ATC 206                                               Gly Val Asn Gln Pro Glu Arg Cys                                              Ser Gly Val Gln Gly Gly Ser Ile                                                         30                  35                                                              40                       - GAC ATC CCC TTC TCC TTC TAT TTC CCC TGG AAG TTG GCC AAG GAT CCA 254                                               Asp Ile Pro Phe Ser Phe Tyr Phe                                              Pro Trp Lys Leu Ala Lys Asp Pro                                                     45                  50                                                              55                           - CAG ATG AGC ATA GCC TGG AAA TGG AAG GAT TTC CAT GGG GAA GTC ATC 302                                               Gln Met Ser Ile Ala Trp Lys Trp                                              Lys Asp Phe His Gly Glu Val Ile                                                 60                  65                                                              70                  75                                                  - TAC AAC TCC TCC CTG CCT TTC                                               ATA CAT GAG CAC TTC AAG GGC CGG                                               CTC 350                                 Tyr Asn Ser Ser Leu Pro Phe Ile His Glu His Phe Lys Gly Arg Leu                                                                       80                                                              85                  90                                                      - ATC CTG AAC TGG ACA CAG GGT                                               CAG AC 376                              Ile Leu Asn Trp Thr Gln Gly Gln                                                            95                                                                - partial human/mouse alignment:                                             hu MGRPLLLPLLPLLLPPAFLQPSGSTGSGPSYLYGVTQPKHLSASMGGSVEIPFSFYYPWE                                                      mo MAQVLLLLSSGCLHAGNSERYNRKNG----                                            --FGVNQPERCSGVQGGSIDIPFSFYFPWK                                                  - hu LATAPDVRISWRRGHFHGQSFYSTRPP                                            SIHKDYVNRLFLNWTEGQKSGFLRISNLQK...       mo LAKDPQMSIAWKWKDFHGEVIYNSSLPFIHEHFKGRLILNWTQGQ...                         __________________________________________________________________________

                                      TABLE 2                                     __________________________________________________________________________    Sequence encoding a protein related to Ig family members, de-                   signated YE01, isolated from an activated monocyte cell library.             See SEQ ID NO: 5 and 6. Signal sequence is indicated. Nucleotide              1247 may be C or T. Sequence analysis suggests YE01 is a member of            the Ig superfamily of receptors, and is closely related to the CD8            family, which contain a V1J-type fold, particularly the Fc receptors          alpha and/or gamma. Because it contains an ITIM (immune receptor              tyrosine-based inhibitory motif)-like motif, the protein may well be          a monocyte version of the KIR proteins, the Killer Inhibitory Re-             ceptors, which send a negative signal to inhibit killer cell                  function. This protein may share similar function in inhibiting               monocyte effector function, e.g., antigen presentation or subse-              quent response initiation. A mouse counterpart is probably encoded            in the EST W55567.                                                           ACCGGTCCGG AATTCCCGGG TCGACCCACG CGTCCGGGAA GCCCCATAGG CAGGAGGCCC                                                     60                                       - CCGGGCAGCA CATCCTGTCT GCTTGTGTCT GCTGCAGAGT TCTGTCCTTG CATTGGTGCG                                                120                                      - CCTCAGGCCA GGCTGCACTG CTGGGACCTG GGCC ATG TCT CCC CAC CCC ACC 172                                                     Met Ser Pro His Pro Thr                                                       -21 -20                             - GCC CTC CTG GGC CTA GTG CTC TGC CTG GCC CAG ACC ATC CAC ACG CAG 220                                               Ala Leu Leu Gly Leu Val Leu Cys                                              Leu Ala Gln Thr Ile His Thr Gln                                                -15                 -10                                                              -5                   1                                                  - GAG GAA GAT CTG CCC AGA CCC                                               TCC ATC TCG GCT GAG CCA GGC ACC                                               GTG 268                                 Glu Glu Asp Leu Pro Arg Pro Ser Ile Ser Ala Glu Pro Gly Thr Val                                                                    5                                                              10                  15                   - ATC CCC CTG GGG AGC CAT GTG ACT TTC GTG TGC CGG GGC CCG GTT GGG 316                                               Ile Pro Leu Gly Ser His Val Thr                                              Phe Val Cys Arg Gly Pro Val Gly                                                         20                  25                                                              30                       - GTT CAA ACA TTC CGC CTG GAG AGG GAG AGT AGA TCC ACA TAC AAT GAT 364                                               Val Gln Thr Phe Arg Leu Glu Arg                                              Glu Ser Arg Ser Thr Tyr Asn Asp                                                     35                  40                                                              45                           - ACT GAA GAT GTG TCT CAA GCT AGT CCA TCT GAG TCA GAG GCC AGA TTC 412                                               Thr Glu Asp Val Ser Gln Ala Ser                                              Pro Ser Glu Ser Glu Ala Arg Phe                                                 50                  55                                                              60                  65                                                  - CGC ATT GAC TCA GTA AGT GAA                                               GGA AAT GCC GGG CCT TAT CGC TGC                                               ATC 460                                 Arg Ile Asp Ser Val Ser Glu Gly Asn Ala Gly Pro Tyr Arg Cys Ile                                                                       70                                                              75                  80                                                      - TAT TAT AAG CCC CCT AAA TGG                                               TCT GAG CAG AGT GAC TAC CTG GAG                                               CTG 508                                 Tyr Tyr Lys Pro Pro Lys Trp Ser Glu Gln Ser Asp Tyr Leu Glu Leu                                                                   85                                                              90                  95                   - CTG GTG AAA GAA ACC TCT GGA GGC CCG GAC TCC CCG GAC ACA GAG CCC 556                                               Leu Val Lys Glu Thr Ser Gly Gly                                              Pro Asp Ser Pro Asp Thr Glu Pro                                                        100                 105                                                             110                       - GGC TCC TCA GCT GGA CCC ACG CAG AGG CCG TCG GAC AAC AGT CAC AAT 604                                               Gly Ser Ser Ala Gly Pro Thr Gln                                              Arg Pro Ser Asp Asn Ser His Asn                                                    115                 120                                                             125                           - GAG CAT GCA CCT GCT TCC CAA GGC CTG AAA GCT GAG CAT CTG TAT ATT 652                                               Glu His Ala Pro Ala Ser Gln Gly                                              Leu Lys Ala Glu His Leu Tyr Ile                                                130                 135                                                             140                 145                                                  - CTC ATC GGG GTC TCA GTG GTC                                               TTC CTC TTC TGT CTC CTC CTC CTG                                               GTC 700                                 Leu Ile Gly Val Ser Val Val Phe Leu Phe Cys Leu Leu Leu Leu Val                                                                      150                                                             155                 160                                                      - CTC TTC TGC CTC CAT CGC CAG                                               AAT CAG ATA AAG CAG GGG CCC CCC                                               AGA 748                                 Leu Phe Cys Leu His Arg Gln Asn Gln Ile Lys Gln Gly Pro Pro Arg                                                                  165                                                              170                 175                  - AGC AAG GAC GAG GAG CAG AAG CCA CAG CAG AGG CCT GAC CTG GCT GTT 796                                               Ser Lys Asp Glu Glu Gln Lys Pro                                              Gln Gln Arg Pro Asp Leu Ala Val                                                        180                 185                                                             190                       - GAT GTT CTA GAG AGG ACA GCA GAC AAG GCC ACA GTC AAT GGA CTT CCT 844                                               Asp Val Leu Glu Arg Thr Ala Asp                                              Lys Ala Thr Val Asn Gly Leu Pro                                                    195                 200                                                             205                           - GAG AAG GAC AGA GAG ACG GAC ACC TCG GCC CTG GCT GCA GGG AGT TCC 892                                               Glu Lys Asp Arg Glu Thr Asp Thr                                              Ser Ala Leu Ala Ala Gly Ser Ser                                                210                 215                                                             220                 225                                                  - CAG GAG GTG ACG TAT GCT CAG                                               CTG GAC CAC TGG GCC CTC ACA CAG                                               AGG 940                                 Gln Glu Val Thr Tyr Ala Gln Leu Asp His Trp Ala Leu Thr Gln Arg                                                                      230                                                             235                 240                                                      - ACA GCC CGG GCT GTG TCC CCA                                               CAG TCC ACA AAG CCC ATG GCC GAG                                               TCC 988                                 Thr Ala Arg Ala Val Ser Pro Gln Ser Thr Lys Pro Met Ala Glu Ser                                                                  245                                                              250                 255                  - ATC ACG TAT GCA GCC GTT GCC AGA CAC TGACCCCATA CCCACCTGGC 1035                                                    Ile Thr Tyr Ala Ala Val Ala Arg                                              His                                             260                 265                                                - CTCTGCACCT GAGGGTAGAA AGTCACTCTA GGAAAAGCCT GAAGCAGCCA TTTGGAAGGC                                                1095                                     - TTCCTGTTGG ATTCCTCTTC ATCTAGAAAG CCAGCCAGGC AGCTGTCCTG GAGACAAGAG                                                1155                                     - CTGGAGACTG GAGGTTTCTA ACCAGCATCC AGAAGGTTCG TTAGCCAGGT GGTCCCTTCT                                                1215                                     - ACAATCGGAC AGCTCCTTGG ACAGACTGTT TCTCAGTTAT TTCCAAAAAC CCAGCTACAG                                                1275                                     - TTCC 1279                                                                   - A similar gene was cloned by expressing cloning using                      a monoclonal antibody DX26, which was raised against the                      immunogen of human NK cell clone NK681.D5, and selected                       for inhibiting killing by NK cell clones of Fc receptor                       bearing target cells (SP2/0). SEQ ID NO: 7 and 8.                           AAAGGCTGCA GAGTTCTGTC CTTGCATTGG TGCGCCTCAG GCCAGGCTGC ACTGCTGGGA                                                     60                                       - CCTGGGCC ATG TCT CCC CAC CCC ACC GCC CTC CTG GGC CTA GTG CTC TGC 110                Met Ser Pro His Pro Thr Ala Leu Leu Gly Leu Val Leu Cys                                                              -21 -20                                                             -15                 -10                  - CTG GCC CAG ACC ATC CAC ACG CAG GAG GAA GAT CTG CCC AGA CCC TCC 158                                               Leu Ala Gln Thr Ile His Thr Gln                                              Glu Glu Asp Leu Pro Arg Pro Ser                                                         -5                   1                                                           5                           - ATC TCG GCT GAG CCA GGC ACC GTG ATC CCC CTG GGG AGC CAT GTG ACT 206                                               Ile Ser Ala Glu Pro Gly Thr Val                                              Ile Pro Leu Gly Ser His Val Thr                                                 10                  15                                                              20                  25                                                  - TTC GTG TGC CGG GGC CCG GTT                                               GGG GTT CAA ACA TTC CGC CTG GAG                                               AGG 254                                 Phe Val Cys Arg Gly Pro Val Gly Val Gln Thr Phe Arg Leu Glu Arg                                                                       30                                                              35                  40                                                      - GAG AGT AGA TCC ACA TAC AAT                                               GAT ACT GAA GAT GTG TCT CAA GCT                                               AGT 302                                 Glu Ser Arg Ser Thr Tyr Asn Asp Thr Glu Asp Val Ser Gln Ala Ser                                                                   45                                                              50                  55                   - CCA TCT GAG TCA GAG GCC AGA TTC CGC ATT GAC TCA GTA AGT GAA GGA 350                                               Pro Ser Glu Ser Glu Ala Arg Phe                                              Arg Ile Asp Ser Val Ser Glu Gly                                                         60                  65                                                              70                       - AAT GCC GGG CCT TAT CGC TGC ATC TAT TAT AAG CCC CCT AAA TGG TCT 398                                               Asn Ala Gly Pro Tyr Arg Cys Ile                                              Tyr Tyr Lys Pro Pro Lys Trp Ser                                                     75                  80                                                              85                           - GAG CAG AGT GAC TAC CTG GAG CTG CTG GTG AAA GAA ACC TCT GGA GGC 446                                               Glu Gln Ser Asp Tyr Leu Glu Leu                                              Leu Val Lys Glu Thr Ser Gly Gly                                                 90                  95                                                             100                 105                                                  - CCG GAC TCC CCG GAC ACA GAG                                               CCC GGC TCC TCA GCT GGA CCC ACG                                               CAG 494                                 Pro Asp Ser Pro Asp Thr Glu Pro Gly Ser Ser Ala Gly Pro Thr Gln                                                                      110                                                             115                 120                                                      - AGG CCG TCG GAC AAC AGT CAC                                               AAT GAG CAT GCA CCT GCT TCC CAA                                               GGC 542                                 Arg Pro Ser Asp Asn Ser His Asn Glu His Ala Pro Ala Ser Gln Gly                                                                  125                                                              130                 135                  - CTG AAA GCT GAG CAT CTG TAT ATT CTC ATC GGG GTC TCA GTG GTC TTC 590                                               Leu Lys Ala Glu His Leu Tyr Ile                                              Leu Ile Gly Val Ser Val Val Phe                                                        140                 145                                                             150                       - CTC TTC TGT CTC CTC CTC CTG GTC CTC TTC TGC CTC CAT CGC CAG AAT 638                                               Leu Phe Cys Leu Leu Leu Leu Val                                              Leu Phe Cys Leu His Arg Gln Asn                                                    155                 160                                                             165                           - CAG ATA AAG CAG GGG CCC CCC AGA AGC AAG GAC GAG GAG CAG AAG CCA 686                                               Gln Ile Lys Gln Gly Pro Pro Arg                                              Ser Lys Asp Glu Glu Gln Lys Pro                                                170                 175                                                             180                 185                                                  - CAG CAG AGG CCT GAC CTG GCT                                               GTT GAT GTT CTA GAG AGG ACA GCA                                               GAC 734                                 Gln Gln Arg Pro Asp Leu Ala Val Asp Val Leu Glu Arg Thr Ala Asp                                                                      190                                                             195                 200                                                      - AAG GCC ACA GTC AAT GGA CTT                                               CCT GAG AAG GAC AGA GAG ACG GAC                                               ACC 782                                 Lys Ala Thr Val Asn Gly Leu Pro Glu Lys Asp Arg Glu Thr Asp Thr                                                                  205                                                              210                 215                  - TCG GCC CTG GCT GCA GGG AGT TCC CAG GAG GTG ACG TAT GCT CAG CTG 830                                               Ser Ala Leu Ala Ala Gly Ser Ser                                              Gln Glu Val Thr Tyr Ala Gln Leu                                                        220                 225                                                             230                       - GAC CAC TGG GCC CTC ACA CAG AGG ACA GCC CGG GCT GTG TCC CCA CAG 878                                               Asp His Trp Ala Leu Thr Gln Arg                                              Thr Ala Arg Ala Val Ser Pro Gln                                                    235                 240                                                             245                           - TCC ACA AAG CCC ATG GCC GAG TCC ATC ACG TAT GCA GCC GTT GCC AGA 926                                               Ser Thr Lys Pro Met Ala Glu Ser                                              Ile Thr Tyr Ala Ala Val Ala Arg                                                250                 255                                                             260                 265                                                  - CAC TGACCCCATA CCCACCTGGC                                                 CTCTGCACCT GAGGGTAGAA AGTCACTCTA                                              979                                     His                                                                            - GGAAAAGCCT GAAGCAGCCA TTTGGAAGGC TTCCTGTTGG ATTCCTCTTC ATCTAGAAAG                                                1039                                     - CCAGCCAGGC AGCTGTCCTG GAGACAAGAG CTGGAGACTG GAGGTTTCTA ACCAGCATCC                                                1099                                     - AGAAGGTTCG TTAGCCAGGT GGTCCCTTCT ACAATCGAGC AGCTCCTTGG ACAGACTGTT                                                1159                                     - TCTCAGTTAT TTCCAGAGAC CCAGCTACAG TTCCCTGGCT GTTTCTAGAG ACCCAGCTTT                                                1219                                     - ATTCACCTGA CTGTTTCCAG AGACCCAGCT AAAGTCACCT GCCTGTTCTA AAGGCCCAGC                                                1279                                     - TACAGCCAAT CAGCCGATTT CCTGAGCAGT GATGCCACCT CCAAGCTTGT CCTAGGTGTC                                                1339                                     - TGCTGTGAAC CTCCAGTGAC CCCAGAGACT TTGCTGTAAT TATCTGCCCT GCTGACCCTA                                                1399                                     - AAGACCTTCC TAGAAGTCAA GAGCTAGCCT TGAGACTGTG CTATACACAC ACAGCTGAGA                                                1459                                     - GCCAAGCCCA GTTCTCTGGG TTGTGCTTTA CTCCACGCAT CAATAAATAA TTTTGAAGGC                                                1519                                     - CTCACATCTG GCAGCCCCAG GCCTGGTCCT GGGTGCATAG GTCTCTCGGA CCCACTCTCT                                                1579                                     - GCCTTCACAG TTGTTCAAAG CTGAGTGAGG GAAACAGGAC TTACGAAAAC GTGTCAGCGT                                                1639                                     - TTTCTTTTTA AAATTTAATT GATCAGGATT GTACGTAAAA AAAAAAAAAA AAAAAAAAAA                                                1699                                     - AAAAAAAAAA AAAAAAAAAA AAAAAAAGG 1728                                        - Nucleic acid and putative amino acid sequence of soluble                   DLAIR-2. The signal sequence runs from about Met (-21) to                     Thr (-1) (SEQ ID NO: 9 and 10).                                             CCACGCGTCC GGGGACCGGG GCC ATG TCT CCA CAC CTC ACT GCT CTC CTG                                                         50                                                                Met Ser Pro His Leu Thr Ala Leu Leu                                           -21 -20                 -15                          - GGC CTA GTG CTC TGC CTG GCC CAG ACC ATC CAC ACG CAG GAG GGG GCC 98                                                Gly Leu Val Leu Cys Leu Ala Gln                                              Thr Ile His Thr Gln Glu Gly Ala                                                        -10                  -5                                                               1                       - CTT CCC AGA CCC TCC ATC TCG GCT GAG CCA GGC ACT GTG ATC TCC CCG 146                                               Leu Pro Arg Pro Ser Ile Ser Ala                                              Glu Pro Gly Thr Val Ile Ser Pro                                                  5                  10                                                              15                  20                                                  - GGG AGC CAT GTG ACT TTC ATG                                               TGC CGG GGC CCG GTT GGG GTT CAA                                               ACA 194                                 Gly Ser His Val Thr Phe Met Cys Arg Gly Pro Val Gly Val Gln Thr                                                                       25                                                              30                  35                                                      - TTC CGC CTG GAG AGG GAG GAT                                               AGA GCC AAG TAC AAA GAT AGT TAT                                               AAT 242                                 Phe Arg Leu Glu Arg Glu Asp Arg Ala Lys Tyr Lys Asp Ser Tyr Asn                                                                   40                                                              45                  50                   - GTG TTT CGA CTT GGT CCA TCT GAG TCA GAG GCC AGA TTC CAC ATT GAC 290                                               Val Phe Arg Leu Gly Pro Ser Glu                                              Ser Glu Ala Arg Phe His Ile Asp                                                         55                  60                                                              65                       - TCA GTA AGT GAA GGA AAT GCC GGG CTT TAT CGC TGC CTC TAT TAT AAG 338                                               Ser Val Ser Glu Gly Asn Ala Gly                                              Leu Tyr Arg Cys Leu Tyr Tyr Lys                                                     70                  75                                                              80                           - CCC CCT GGA TGG TCT GAG CAC AGT GAC TTC CTG GAG CTG CTG GTG AAA 386                                               Pro Pro Gly Trp Ser Glu His Ser                                              Asp Phe Leu Glu Leu Leu Val Lys                                                 85                  90                                                              95                 100                                                  - GGG ACT GTG CCA GGC ACT GAA                                               GCC TCC GGA TTT GAT GCA CCA 428                                                Gly Thr Val Pro Gly Thr Glu Ala                                              Ser Gly Phe Asp Ala Pro                                 105                 110                                        - TGAATGAGGA GAAATGGCCT CCCGTCTTGT GAACTTCAAT GGGGAGAAAT AATTAGAATG                                                488                                      - AGCAATAGAA ATGCACAGAT GCCTATACAT ACATATACAA ATAAAAAGAT ACGATTCGCA                                                548                                      - AAAAAAAAAA AAAAAAGGGC 568                                                __________________________________________________________________________

                                      TABLE 3                                     __________________________________________________________________________    Human KTE03 sequences, e.g., alternative splicing, encoding related            proteins with homology to several NK KIR surface molecules, and to            the Fc receptors gamma and alpha. YYB01 coding sequence appears to            run from about 81 to 1397. The message appears to be IL-10                    upregulated. See SEQ ID NO: 11 and 12. Because of significant                 identity of sequence which ends at specific locations, it appears             that there may be splice junctions around nucleotide 36, 1264, and            1587. The YYB04 sequence provided below indicates that certain                insertions of sequence lead to a frameshift and alternative carboxy           terminal sequence. Moreover, certain peculiar differences in                  sequence suggest either sequencing errors, or a mechanism of                  variability generated by a mechanism perhaps analogous to                     hypervariable region combinations.                                           GTCGACCCAC GCGTCCGCCT CTGTCCTGCC AGCACCGAGG GCTCATCCAT CCACAGAGCA                                                     60                                       - GTGCAGTGGG AGGAGACGCC ATG ACC CCC ATC CTC ACG GTC CTG ATC TGT 110                                                                       Met Thr                                                Pro Ile Leu Thr Val Leu Ile Cys                                                                        1                                                           5                   10                                                   - CTC GGG CTG AGC CTG GAC CCC                                               AGG ACC CAC GTG CAG GCA GGG CCC                                               CTC 158                                 Leu Gly Leu Ser Leu Asp Pro Arg Thr His Val Gln Ala Gly Pro Leu                                                                       15                                                               20                   25                                                    - CCC AAG CCC ACC CTC TGG GCT                                               GAG CCA GGC TCT GTG ATC ACC CAA                                               GGG 206                                 Pro Lys Pro Thr Leu Trp Ala Glu Pro Gly Ser Val Ile Thr Gln Gly                                                                   30                                                               35                   40                 - AGT CCT GTG ACC CTC AGG TGT CAG GGG AGC CTG GAG ACG CAG GAG TAC 254                                               Ser Pro Val Thr Leu Arg Cys Gln                                              Gly Ser Leu Glu Thr Gln Glu Tyr                                                         45                   50                                                               55                     - CAT CTA TAT AGA GAA AAG AAA ACA GCA CTC TGG ATT ACA CGG ATC CCA 302                                               His Leu Tyr Arg Glu Lys Lys Thr                                              Ala Leu Trp Ile Thr Arg Ile Pro                                                     60                   65                                                               70                         - CAG GAG CTT GTG AAG AAG GGC CAG TTC CCC ATC CTA TCC ATC ACC TGG 350                                               Gln Glu Leu Val Lys Lys Gly Gln                                              Phe Pro Ile Leu Ser Ile Thr Trp                                                 75                   80                                                               85                   90        - GAA CAT GCA GGG CGG TAT TGC TGT ATC TAT GGC AGC CAC ACT GCA GGC 398                                               Glu His Ala Gly Arg Tyr Cys Cys                                              Ile Tyr Gly Ser His Thr Ala Gly                                                                 95                                                             100                 105                                                      - CTC TCA GAG AGC AGT GAC CCC                                               CTG GAG CTG GTG GTG ACA GGA GCC                                               TAC 446                                 Leu Ser Glu Ser Ser Asp Pro Leu Glu Leu Val Val Thr Gly Ala Tyr                                                                  110                                                              115                 120                  - AGC AAA CCC ACC CTC TCA GCT CTG CCC AGC CCT GTG GTG ACC TCA GGA 494                                               Ser Lys Pro Thr Leu Ser Ala Leu                                              Pro Ser Pro Val Val Thr Ser Gly                                                        125                 130                                                             135                       - GGG AAT GTG ACC ATC CAG TGT GAC TCA CAG GTG GCA TTT GAT GGC TTC 542                                               Gly Asn Val Thr Ile Gln Cys Asp                                              Ser Gln Val Ala Phe Asp Gly Phe                                                    140                 145                                                             150                           - ATT CTG TGT AAG GAA GGA GAA GAT GAA CAC CCA CAA TGC CTG AAC TCC 590                                               Ile Leu Cys Lys Glu Gly Glu Asp                                              Glu His Pro Gln Cys Leu Asn Ser                                                155                 160                                                             165                 170                                                  - CAT TCC CAT GCC CGT GGG TCA                                               TCC CGG GCC ATC TTC TCC GTG GGC                                               CCC 638                                 His Ser His Ala Arg Gly Ser Ser Arg Ala Ile Phe Ser Val Gly Pro                                                                      175                                                             180                 185                                                      - GTG AGC CCA AGT CGC AGG TGG                                               TCG TAC AGG TGC TAT GGT TAT GAC                                               TCG 686                                 Val Ser Pro Ser Arg Arg Trp Ser Tyr Arg Cys Tyr Gly Tyr Asp Ser                                                                  190                                                              195                 200                  - CGC GCT CCC TAT GTG TGG TCT CTA CCC AGT GAT CTC CTG GGG CTC CTG 734                                               Arg Ala Pro Tyr Val Trp Ser Leu                                              Pro Ser Asp Leu Leu Gly Leu Leu                                                        205                 210                                                             215                       - GTC CCA GGT GTT TCT AAG AAG CCA TCA CTC TCA GTG CAG CCG GGT CCT 782                                               Val Pro Gly Val Ser Lys Lys Pro                                              Ser Leu Ser Val Gln Pro Gly Pro                                                    220                 225                                                             230                           - GTC GTG GCC CCT GGG GAG AAG CTG ACC TTC CAG TGT GGC TCT GAT GCC 830                                               Val Val Ala Pro Gly Glu Lys Leu                                              Thr Phe Gln Cys Gly Ser Asp Ala                                                235                 240                                                             245                 250                                                  - GGC TAC GAC AGA TTT GTT CTG                                               TAC AAG GAG TGG GGA CGT GAC TTC                                               CTC 878                                 Gly Tyr Asp Arg Phe Val Leu Tyr Lys Glu Trp Gly Arg Asp Phe Leu                                                                      255                                                             260                 265                                                      - CAG CGC CCT GGC CGG CAG CCC                                               CAG GCT GGG CTC TCC CAG GCC AAC                                               TTC 926                                 Gln Arg Pro Gly Arg Gln Pro Gln Ala Gly Leu Ser Gln Ala Asn Phe                                                                  270                                                              275                 280                  - ACC CTG GGC CCT GTG AGC CGC TCC TAC GGG GGC CAG TAC ACA TGC TCC 974                                               Thr Leu Gly Pro Val Ser Arg Ser                                              Tyr Gly Gly Gln Tyr Thr Cys Ser                                                        285                 290                                                             295                       - GGT GCA TAC AAC CTC TCC TCC GAG TGG TCG GCC CCC AGC GAC CCC CTG 1022       Gly Ala Tyr Asn Leu Ser Ser Glu Trp Ser Ala Pro Ser Asp Pro Leu                                                          300                 305                                                             310                           - GAC ATC CTG ATC ACA GGA CAG ATC CGT GCC AGA CCC TTC CTC TCC GTG 1070       Asp Ile Leu Ile Thr Gly Gln Ile Arg Ala Arg Pro Phe Leu Ser Val                                                      315                 320                                                             325                 330                                                  - CGG CCG GGC CCC ACA GTG GCC                                               TCA GGA GAG AAC GTG ACC CTG CTG                                               TGT 1118                                Arg Pro Gly Pro Thr Val Ala Ser Gly Glu Asn Val Thr Leu Leu Cys                                                                      335                                                             340                 345                                                      - CAG TCA CAG GGA GGG ATG CAC                                               ACT TTC CTT TTG ACC AAG GAG GGG                                               GCA 1166                                Gln Ser Gln Gly Gly Met His Thr Phe Leu Leu Thr Lys Glu Gly Ala                                                                  350                                                              355                 360                  - GCT GAT TCC CCG CTG CGT CTA AAA TCA AAG CGC CAA TCT CAT AAG TAC 1214       Ala Asp Ser Pro Leu Arg Leu Lys Ser Lys Arg Gln Ser His Lys Tyr                                                              365                 370                                                             375                       - CAG GCT GAA TTC CCC ATG AGT CCT GTG ACC TCG GCC CAC GCG GGG ACC 1262       Gln Ala Glu Phe Pro Met Ser Pro Val Thr Ser Ala His Ala Gly Thr                                                          380                 385                                                             390                           - TAC AGG TGC TAC GGC TCA CTC AGC TCC AAC CCC TAC CTG CTG ACT CAC 1310       Tyr Arg Cys Tyr Gly Ser Leu Ser Ser Asn Pro Tyr Leu Leu Thr His                                                      395                 400                                                             405                 410                                                  - CCC AGT GAC CCC CTG GAG CTC                                               GTG GTC TCA GGA GCA GCT GAG ACC                                               CTC 1358                                Pro Ser Asp Pro Leu Glu Leu Val Val Ser Gly Ala Ala Glu Thr Leu                                                                      415                                                             420                 425                                                      - AGC CCA CCA CAA AAC AAG TCC                                               GAC TCC AAG GCT GGT GAG TGAGGAGATG                                             1407                                   Ser Pro Pro Gln Asn Lys Ser Asp Ser Lys Ala Gly Glu                                       430                 435                                            - CTTGCCGTGA TGACGCTGGG CACAGAGGGT CAGGTCCTGT CAAGAGGAGC TGGGTGTCCT                                                1467                                     - GGGTGGACAT TTGAAGAATT ATATTCATTC CAACTTGAAG AATTATTCAA CACCTTTAAC                                                1527                                     - AATGTATATG TGAAGTACTT TATTCTTTCA TATTTTAAAA ATAAAAGATA ATTATCCATG                                                1587                                     - AAAAAAAAAA AAAAAAAAAA AAAGGGCGGC CGC 1620                                   -                                                                          YYB04: Related to YYB01, apparently through alternative splicing                from the same or a very highly related gene. The coding region runs          from about 191 to 1493, but the initiation methionine may actually            be at the numbered Met at 18. See SEQ ID NO: 13 and 14. Another               transcript was isolated which contains evidence for existence of an           insert of sequence TGCTACGGCT CACTCCAACTC CGACCCCTAC CTGCTGTCTC               ACCCCAGTGA GCCCCTGGAG CTCGTGGTCT CAGG between residues 1426 and               1427, which changes the downstream reading frame of the subsequent            sequence, to encode, from residue 413, CYG SLNSD PYLLS HPSEP LELVV            SGPSM GSSPP PTGPI STPAG PEDQP LTPTG SDPQS GLGRH LGVVI GILVA VVLLL             LLLLL LFLIL RHRRQ GKHWT STQRK ADFQH PAGAV GPEPT DRGLQ WRSSP AADAQ             EENLY AAVKD TQPED GVEMD TRAAA SEAPQ DVTYA QLHSL TLRRK ATEPP PSQER             EPPAE TLAIH. (SEQ ID NO: 15 and 16.) This alternative sequence                contains a transmembrane segment from about 478 to 500.                      GTCGACCCAC GCGTCCGGTC AACTTTTCTT CCCCTACTTC CCTGCATTTC TCCTCTGTGC                                                     60                                       - TCACTGCCAC ACGCAGCTCA ACCTGGACGG CACAGCCAGA TGCGAGATGC GTCTCTGCTG                                                120                                      - ATCTGAGTCT GCCTGCAGCA TGGACCTGGG TCTTCCCTGA AGCATCTCCA GGGCTGGAGG                                                180                                      - GACGACTGCC ATG CAC CGA GGG CTC ATC CAT CCG CAG AGC AGG GCA GTG 229                                                           Met His Arg Gly Leu                                               Ile His Pro Gln Ser Arg Ala Val                                                             1               5                                                               10                       - GGA GGA GAC GCC ATG ACC CCC ATC GTC ACA GTC CTG ATC TGT CTC GGG 277                                               Gly Gly Asp Ala Met Thr Pro Ile                                              Val Thr Val Leu Ile Cys Leu Gly                                                     15                   20                                                               25                         - CTG AGT CTG GGC CCC AGG ACC CAC GTG CAG ACA GGG ACC ATC CCC AAG 325                                               Leu Ser Leu Gly Pro Arg Thr His                                              Val Gln Thr Gly Thr Ile Pro Lys                                                 30                   35                                                               40                   45        - CCC ACC CTG TGG GCT GAG CCA GAC TCT GTG ATC ACC CAG GGG AGT CCC 373                                               Pro Thr Leu Trp Ala Glu Pro Asp                                              Ser Val Ile Thr Gln Gly Ser Pro                                                                 50                                                               55                   60                                                    - GTC ACC CTC AGT TGT CAG GGG                                               AGC CTT GAA GCC CAG GAG TAC CGT                                               CTA 421                                 Val Thr Leu Ser Cys Gln Gly Ser Leu Glu Ala Gln Glu Tyr Arg Leu                                                                   65                                                               70                   75                 - TAT AGG GAG AAA AAA TCA GCA TCT TGG ATT ACA CGG ATA CGA CCA GAG 469                                               Tyr Arg Glu Lys Lys Ser Ala Ser                                              Trp Ile Thr Arg Ile Arg Pro Glu                                                         80                   85                                                               90                     - CTT GTG AAG AAC GGC CAG TTC CAC ATC CCA TCC ATC ACC TGG GAA CAC 517                                               Leu Val Lys Asn Gly Gln Phe His                                              Ile Pro Ser Ile Thr Trp Glu His                                                     95                 100                                                             105                           - ACA GGG CGA TAT GGC TGT CAG TAT TAC AGC CGC GCT CGG TGG TCT GAG 565                                               Thr Gly Arg Tyr Gly Cys Gln Tyr                                              Tyr Ser Arg Ala Arg Trp Ser Glu                                                110                  115                                                             120                 125                                                 - CTC AGT GAC CCC CTG GTG CTG                                               GTG ATG ACA GGA GCC TAC CCA AAA                                               CCC 613                                 Leu Ser Asp Pro Leu Val Leu Val Met Thr Gly Ala Tyr Pro Lys Pro                                                                      130                                                             135                 140                                                      - ACC CTC TCA GCC CAG CCC AGC                                               CCT GTG GTG ACC TCA GGA GGA AGG                                               GTG 661                                 Thr Leu Ser Ala Gln Pro Ser Pro Val Val Thr Ser Gly Gly Arg Val                                                                  145                                                              150                 155                  - ACC CTC CAG TGT GAG TCA CAG GTG GCA TTT GGC GGC TTC ATT CTG TGT 709                                               Thr Leu Gln Cys Glu Ser Gln Val                                              Ala Phe Gly Gly Phe Ile Leu Cys                                                        160                 165                                                             170                       - AAG GAA GGA GAA GAT GAA CAC CCA CAA TGC CTG AAC TCC CAG CCC CAT 757                                               Lys Glu Gly Glu Asp Glu His Pro                                              Gln Cys Leu Asn Ser Gln Pro His                                                    175                 180                                                             185                           - GCC CGT GGG TCG TCC CGC GCC ATC TTC TCC GTG GGC CCC GTG AGC CCG 805                                               Ala Arg Gly Ser Ser Arg Ala Ile                                              Phe Ser Val Gly Pro Val Ser Pro                                                190                 195                                                             200                 205                                                  - AAT CGC AGG TGG TCG CAC AGG                                               TGC TAT GGT TAT GAC TTG AAC TCT                                               CCC 853                                 Asn Arg Arg Trp Ser His Arg Cys Tyr Gly Tyr Asp Leu Asn Ser Pro                                                                      210                                                             215                 220                                                      - TAT GTG TGG TCT TCA CCC AGT                                               GAT CTC CTG GAG CTC CTG GTC CCA                                               GGT 901                                 Tyr Val Trp Ser Ser Pro Ser Asp Leu Leu Glu Leu Leu Val Pro Gly                                                                  225                                                              230                 235                  - GTT TCT AAG AAG CCA TCA CTC TCA GTG CAG CCG GGT CCT GTC GTG GCC 949                                               Val Ser Lys Lys Pro Ser Leu Ser                                              Val Gln Pro Gly Pro Val Val Ala                                                        240                 245                                                             250                       - CCT GGG GAA AGC CTG ACC CTC CAG TGT GTC TCT GAT GTC GGC TAT GAC 997                                               Pro Gly Glu Ser Leu Thr Leu Gln                                              Cys Val Ser Asp Val Gly Tyr Asp                                                    255                 260                                                             265                           - AGA TTT GTT CTG TAC AAG GAG GGG GAA CGT GAC CTT CGC CAG CTC CCT 1045       Arg Phe Val Leu Tyr Lys Glu Gly Glu Arg Asp Leu Arg Gln Leu Pro                                                      270                 275                                                             280                 285                                                  - GGC CGG CAG CCC CAG GCT GGG                                               CTC TCC CAG GCC AAC TTC ACC CTG                                               GGC 1093                                Gly Arg Gln Pro Gln Ala Gly Leu Ser Gln Ala Asn Phe Thr Leu Gly                                                                      290                                                             295                 300                                                      - CCT GTG AGC CGC TCC TAC GGG                                               GGC CAG TAC AGA TGC TAC GGT GCA                                               TAC 1141                                Pro Val Ser Arg Ser Tyr Gly Gly Gln Tyr Arg Cys Tyr Gly Ala Tyr                                                                  305                                                              310                 315                  - AAC CTC TCC TCC GAG TGG TCG GCC CCC AGC GAC CCC CTG GAC ATC CTG 1189       Asn Leu Ser Ser Glu Trp Ser Ala Pro Ser Asp Pro Leu Asp Ile Leu                                                              320                 325                                                             330                       - ATC ACA GGA CAG ATC CAT GGC ACA CCC TTC ATC TCA GTG CAG CCA GGC 1237       Ile Thr Gly Gln Ile His Gly Thr Pro Phe Ile Ser Val Gln Pro Gly                                                          335                 340                                                             345                           - CCC ACA GTG GCC TCA GGA GAG AAC GTG ACC CTG CTG TGT CAG TCA TGG 1285       Pro Thr Val Ala Ser Gly Glu Asn Val Thr Leu Leu Cys Gln Ser Trp                                                      350                 355                                                             360                 365                                                  - CGG CAG TTC CAC ACT TTC CTT                                               CTG ACC AAG GCG GGA GCA GCT GAT                                               GCC 1333                                Arg Gln Phe His Thr Phe Leu Leu Thr Lys Ala Gly Ala Ala Asp Ala                                                                      370                                                             375                 380                                                      - CCA CTC CGT CTA AGA TCA ATA                                               CAC GAA TAT CCT AAG TAC CAG GCT                                               GAA 1381                                Pro Leu Arg Leu Arg Ser Ile His Glu Tyr Pro Lys Tyr Gln Ala Glu                                                                  385                                                              390                 395                  - TTC CCC ATG AGT CCT GTG ACC TCA GCC CAC GCG GGG ACC TAC AGG ACC 1429       Phe Pro Met Ser Pro Val Thr Ser Ala His Ala Gly Thr Tyr Arg Thr                                                              400                 405                                                             410                       - CTC CAT GGG TTC CAG CCC CCC ACC CAC CGG TCC CAT CTC CAC ACC TGC 1477       Leu His Gly Phe Gln Pro Pro Thr His Arg Ser His Leu His Thr Cys                                                          415                 420                                                             425                           - AGG CCC TGAGGACCAG CCCCTCACCC CCACTGGGTC GGATCCCCAA AGTGGTCTGG 1533                                               Arg Pro                                430                                                                            - GAAGGCACCT GGGGGTTGTG ATCGGCATCT TGGTGGCCGT CGTCCTACTG CTCCTCCTCC                                                1593                                     - TCCTCCTCCT CTTCCTCATC CTCCGACATC GACGTCAGGG CAAACACTGG ACATCGACCC                                                1653                                     - AGAGAAAGGC TGATTTCCAA CATCCTGCAG GGGCTGTGGG GCCAGAGCCC ACAGACAGAG                                                1713                                     - GCCTGCAGTG GAGGTCCAGC CCAGCTGCCG ACGCCCAGGA AGAAAACCTC TATGCTGCCG                                                1773                                     - TGAAGGACAC ACAGCCTGAA GATGGGGTGG AGATGGACAC TCGGGCTGCT GCATCTGAAG                                                1833                                     - CCCCCCAGGA TGTGACCTAC GCCCAGCTGC ACAGCTTGAC CCTCAGACGG AAGGCAACTG                                                1893                                     - AGCCTCCTCC ATCCCAGGAA AGGGAACCTC CAGCTGAGCC CAGCATTTAC GCCACCCTGG                                                1953                                     - CCATCCACTA GCCCGGAGGG TACGCAGACT CCACACTCAG TAGAAGGAGA CTCAGGACTG                                                2013                                     - CTGAAGGCAC GGGAGCTGCC CCCAGTGGAC ACCAATGAAC CCCAGTCAGC CTGGACCCCT                                                2073                                     - AACAAAGACC ATGAGGAGAT GCTGGGAACT TTGGGACTCA CTTGATTCTG CAGTGGAAAT                                                2133                                     - AACTAATATC CCTACATTTT TTAATTAAAG CAACAGACTT CTCAATAATC AATGAGTTAA                                                2193                                     - CCGA 2197                                                                   -                                                                          A KTE03 embodiment designated KLM63 (SEQ ID NO: 17 and 18):                   AAAGAAGTCA ACTTTTCTTC CCCTACTTCC CTGCATTTCT CCTCTGTGCT CACTGCCACA                                                     60                                       - CGCAGCTCAA CCTGGACGGC ACAGCCAGAT GCGAGATGCG TCTCTGCTGA TCTGAGTCTG                                                120                                      - CCTGCAGCAT GGACCTGGGT CTTCCCTGAA GCATCTCCAG GGCTGGAGGG ACGACTGCC 179        - ATG CAC CGA GGG CTC ATC CAT CCG CAG AGC AGG GCA GTG GGA GGA GAC 227                                               Met His Arg Gly Leu Ile His Pro                                              Gln Ser Arg Ala Val Gly Gly Asp                                                  1               5                                                               10                   15                                                    - GCC ATG ACC CCC ATC GTC ACA                                               GTC CTG ATC TGT CTC GGG CTG AGT                                               CTG 275                                 Ala Met Thr Pro Ile Val Thr Val Leu Ile Cys Leu Gly Leu Ser Leu                                                                   20                                                               25                   30                 - GGC CCC AGG ACC CAC GTG CAG ACA GGG ACC ATC CCC AAG CCC ACC CTG 323                                               Gly Pro Arg Thr His Val Gln Thr                                              Gly Thr Ile Pro Lys Pro Thr Leu                                                         35                   40                                                               45                     - TGG GCT GAG CCA GAC TCT GTG ATC ACC CAG GGG AGT CCC GTC ACC CTC 371                                               Trp Ala Glu Pro Asp Ser Val Ile                                              Thr Gln Gly Ser Pro Val Thr Leu                                                     50                   55                                                               60                         - AGT TGT CAG GGG AGC CTT GAA GCC CAG GAG TAC CGT CTA TAT AGG GAG 419                                               Ser Cys Gln Gly Ser Leu Glu Ala                                              Gln Glu Tyr Arg Leu Tyr Arg Glu                                                 65                   70                                                               75                   80        - AAA AAA TCA GCA TCT TGG ATT ACA CGG ATA CGA CCA GAG CTT GTG AAG 467                                               Lys Lys Ser Ala Ser Trp Ile Thr                                              Arg Ile Arg Pro Glu Leu Val Lys                                                                 85                                                               90                   95                                                    - AAC GGC CAG TTC CAC ATC CCA                                               TCC ATC ACC TGG GAA CAC ACA GGG                                               CGA 515                                 Asn Gly Gln Phe His Ile Pro Ser Ile Thr Trp Glu His Thr Gly Arg                                                                  100                                                              105                 110                  - TAT GGC TGT CAG TAT TAC AGC CGC GCT CGG TGG TCT GAG CTC AGT GAC 563                                               Tyr Gly Cys Gln Tyr Tyr Ser Arg                                              Ala Arg Trp Ser Glu Leu Ser Asp                                                        115                 120                                                             125                       - CCC CTG GTG CTG GTG ATG ACA GGA GCC TAC CCA AAA CCC ACC CTC TCA 611                                               Pro Leu Val Leu Val Met Thr Gly                                              Ala Tyr Pro Lys Pro Thr Leu Ser                                                    130                 135                                                             140                           - GCC CAG CCC AGC CCT GTG GTG ACC TCA GGA GGA AGG GTG ACC CTC CAG 659                                               Ala Gln Pro Ser Pro Val Val Thr                                              Ser Gly Gly Arg Val Thr Leu Gln                                                145                 150                                                             155                 160                                                  - TGT GAG TCA CAG GTG GCA TTT                                               GGC GGC TTC ATT CTG TGT AAG GAA                                               GGA 707                                 Cys Glu Ser Gln Val Ala Phe Gly Gly Phe Ile Leu Cys Lys Glu Gly                                                                      165                                                             170                 175                                                      - GAA GAT GAA CAC CCA CAA TGC                                               CTG AAC TCC CAG CCC CAT GCC CGT                                               GGG 755                                 Glu Asp Glu His Pro Gln Cys Leu Asn Ser Gln Pro His Ala Arg Gly                                                                  180                                                              185                 190                  - TCG TCC CGC GCC ATC TTC TCC GTG GGC CCC GTG AGC CCG AAT CGC AGG 803                                               Ser Ser Arg Ala Ile Phe Ser Val                                              Gly Pro Val Ser Pro Asn Arg Arg                                                        195                 200                                                             205                       - TGG TCG CAC AGG TGC TAT GGT TAT GAC TTG AAC TCT CCC TAT GTG TGG 851                                               Trp Ser His Arg Cys Tyr Gly Tyr                                              Asp Leu Asn Ser Pro Tyr Val Trp                                                    210                 215                                                             220                           - TCT TCA CCC AGT GAT CTC CTG GAG CTC CTG GTC CCA GGT GTT TCT AAG 899                                               Ser Ser Pro Ser Asp Leu Leu Glu                                              Leu Leu Val Pro Gly Val Ser Lys                                                225                 230                                                             235                 240                                                  - AAG CCA TCA CTC TCA GTG CAG                                               CCG GGT CCT GTC GTG GCC CCT GGG                                               GAA 947                                 Lys Pro Ser Leu Ser Val Gln Pro Gly Pro Val Val Ala Pro Gly Glu                                                                      245                                                             250                 255                                                      - AGC CTG ACC CTC CAG TGT GTC                                               TCT GAT GTC GGC TAT GAC AGA TTT                                               GTT 995                                 Ser Leu Thr Leu Gln Cys Val Ser Asp Val Gly Tyr Asp Arg Phe Val                                                                  260                                                              265                 270                  - CTG TAC AAG GAG GGG GAA CGT GAC CTT CGC CAG CTC CCT GGC CGG CAG 1043       Leu Tyr Lys Glu Gly Glu Arg Asp Leu Arg Gln Leu Pro Gly Arg Gln                                                              275                 280                                                             285                       - CCC CAG GCT GGG CTC TCC CAG GCC AAC TTC ACC CTG GGC CCT GTG AGC 1091       Pro Gln Ala Gly Leu Ser Gln Ala Asn Phe Thr Leu Gly Pro Val Ser                                                          290                 295                                                             300                           - CGC TCC TAC GGG GGC CAG TAC AGA TGC TAC GGT GCA TAC AAC CTC TCC 1139       Arg Ser Tyr Gly Gly Gln Tyr Arg Cys Tyr Gly Ala Tyr Asn Leu Ser                                                      305                 310                                                             315                 320                                                  - TCC GAG TGG TCG GCC CCC AGC                                               GAC CCC CTG GAC ATC CTG ATC ACA                                               GGA 1187                                Ser Glu Trp Ser Ala Pro Ser Asp Pro Leu Asp Ile Leu Ile Thr Gly                                                                      325                                                             330                 335                                                      - CAG ATC CAT GGC ACA CCC TTC                                               ATC TCA GTG CAG CCA GGC CCC ACA                                               GTG 1235                                Gln Ile His Gly Thr Pro Phe Ile Ser Val Gln Pro Gly Pro Thr Val                                                                  340                                                              345                 350                  - GCC TCA GGA GAG AAC GTG ACC CTG CTG TGT CAG TCA TGG CGG CAG TTC 1283       Ala Ser Gly Glu Asn Val Thr Leu Leu Cys Gln Ser Trp Arg Gln Phe                                                              355                 360                                                             365                       - CAC ACT TTC CTT CTG ACC AAG GCG GGA GCA GCT GAT GCC CCA CTC CGT 1331       His Thr Phe Leu Leu Thr Lys Ala Gly Ala Ala Asp Ala Pro Leu Arg                                                          370                 375                                                             380                           - CTA AGA TCA ATA CAC GAA TAT CCT AAG TAC CAG GCT GAA TTC CCC ATG 1379       Leu Arg Ser Ile His Glu Tyr Pro Lys Tyr Gln Ala Glu Phe Pro Met                                                      385                 390                                                             395                 400                                                  - AGT CCC GTG ACC TCA GCC CAC                                               GCG GGG ACC TAC AGG TGC TAC GGC                                               TCA 1427                                Ser Pro Val Thr Ser Ala His Ala Gly Thr Tyr Arg Cys Tyr Gly Ser                                                                      405                                                             410                 415                                                      - CTC AAC TCC GAC CCC TAC CTG                                               CTG TCT CAC CCC AGT GAG CCC CTG                                               GAG 1475                                Leu Asn Ser Asp Pro Tyr Leu Leu Ser His Pro Ser Glu Pro Leu Glu                                                                  420                                                              425                 430                  - CTC GTG GTC TCA GGA CCC TCC ATG GGT TCC AGC CCC CCA CCC ACC GGT 1523       Leu Val Val Ser Gly Pro Ser Met Gly Ser Ser Pro Pro Pro Thr Gly                                                              435                 440                                                             445                       - CCC ATC TCC ACA CCT GCA GGC CCT GAG GAC CAG CCC CTC ACC CCC ACT 1571       Pro Ile Ser Thr Pro Ala Gly Pro Glu Asp Gln Pro Leu Thr Pro Thr                                                          450                 455                                                             460                           - GGG TCG GAT CCC CAA AGT GGT CTG GGA AGG CAC CTG GGG GTT GTG ATC 1619       Gly Ser Asp Pro Gln Ser Gly Leu Gly Arg His Leu Gly Val Val Ile                                                      465                 470                                                             475                 480                                                  - GGC ATC TTG GTG GCC GTC GTC                                               CTA CTG CTC CTC CTC CTC CTC CTC                                               CTC 1667                                Gly Ile Leu Val Ala Val Val Leu Leu Leu Leu Leu Leu Leu Leu Leu                                                                      485                                                             490                 495                                                      - TTC CTC ATC CTC CGA CAT CGA                                               CGT CAG GGC AAA CAC TGG ACA TCG                                               ACC 1715                                Phe Leu Ile Leu Arg His Arg Arg Gln Gly Lys His Trp Thr Ser Thr                                                                  500                                                              505                 510                  - CAG AGA AAG GCT GAT TTC CAA CAT CCT GCA GGG GCT GTG GGG CCA GAG 1763       Gln Arg Lys Ala Asp Phe Gln His Pro Ala Gly Ala Val Gly Pro Glu                                                              515                 520                                                             525                       - CCC ACA GAC AGA GGC CTG CAG TGG AGG TCC AGC CCA GCT GCC GAC GCC 1811       Pro Thr Asp Arg Gly Leu Gln Trp Arg Ser Ser Pro Ala Ala Asp Ala                                                          530                 535                                                             540                           - CAG GAA GAA AAC CTC TAT GCT GCC GTG AAG GAC ACA CAG CCT GAA GAT 1859       Gln Glu Glu Asn Leu Tyr Ala Ala Val Lys Asp Thr Gln Pro Glu Asp                                                      545                  550                                                             555                 560                                                 - GGG GTG GAG ATG GAC ACT CGG                                               GCT GCT GCA TCT GAA GCC CCC CAG                                               GAT 1907                                Gly Val Glu Met Asp Thr Arg Ala Ala Ala Ser Glu Ala Pro Gln Asp                                                                      565                                                             570                 575                                                      - GTG ACC TAC GCC CAG CTG CAC                                               AGC TTG ACC CTC AGA CGG AAG GCA                                               ACT 1955                                Val Thr Tyr Ala Gln Leu His Ser Leu Thr Leu Arg Arg Lys Ala Thr                                                                  580                                                              585                 590                  - GAG CCT CCT CCA TCC CAG GAA AGG GAA CCT CCA GCT GAG CCC AGC ATC 2003       Glu Pro Pro Pro Ser Gln Glu Arg Glu Pro Pro Ala Glu Pro Ser Ile                                                              595                 600                                                             605                       - TAC GCC ACC CTG GCC ATC CAC TAGCCCGGAG GGTACGCAGA CTCCACACTC 2054                                                 Tyr Ala Thr Leu Ala Ile His                                                       610                 615                                                    - AGTAGAAGGA GACTCAGGAC                                                     TGCTGAAGGC ACGGGAGCTG CCCCCAGTGG                                              ACACCAATGA 2114                          - ACCCCAGTCA GCCTGGACCC CTAACAAAGA CCATGAGGAG ATGCTGGGAA CTTTGGGACT                                                2174                                     - CACTTGATTC TGCAGTCGAA ATAACTAATA TCCCTACATT TTTTAATTAA AGCAACAGAC                                                2234                                     - TTCTCAATAA TCAATGAGTT AACCGAGAAA ACTAAAATCA GAAGTAAGAA TGTGCTTTAA                                                2294                                     - ACTGAATCAC AATATAAATA TTACACATCA CACAATGAAA TTGAAAAAGT ACAAACCACA                                                2354                                     - AATGAAAAAA GTAGAAACGA AAAAAAAAAA AAAA 2388                                  -                                                                          A KTE03 embodiment designated KLM66 (SEQ IDS NO: 19 and 20):                  GTCAACTTTT CTTCCCCTAC TTCCCTGCAT TTCTCCTCTG TGCTCACTGC CACACGCAGC                                                     60                                       - TCAACCTGGA CGGCACAGCC AGATGCGAGA TGCGTCTCTG CTGATCTGAG TCTGCCTGCA                                                120                                      - GCATGGACCT GGGTCTTCCC TGAAGCATCT CCAGGGCTGG AGGGACGACT GCC ATG 176                                               Met                                                                                                       1                                                   - CAC CGA GGG CTC ATC CAT CCG                                               CAG AGC AGG GCA GTG GGA GGA GAC                                               GCC 224                                 His Arg Gly Leu Ile His Pro Gln Ser Arg Ala Val Gly Gly Asp Ala                                                                    5                                                               10                   15                 - ATG ACC CCC ATC GTC ACA GTC CTG ATC TGT CTC GGG CTG AGT CTG GGC 272                                               Met Thr Pro Ile Val Thr Val Leu                                              Ile Cys Leu Gly Leu Ser Leu Gly                                                         20                  25                                                                30                     - CCC AGG ACC CAC GTG CAG ACA GGG ACC ATC CCC AAG CCC ACC CTG TGG 320                                               Pro Arg Thr His Val Gln Thr Gly                                              Thr Ile Pro Lys Pro Thr Leu Trp                                                     35                   40                                                               45                         - GCT GAG CCA GAC TCT GTG ATC ACC CAG GGG AGT CCC GTC ACC CTC AGT 368                                               Ala Glu Pro Asp Ser Val Ile Thr                                              Gln Gly Ser Pro Val Thr Leu Ser                                                 50                   55                                                               60                   65        - TGT CAG GGG AGC CTT GAA GCC CAG GAG TAC CGT CTA TAT AGG GAG AAA 416                                               Cys Gln Gly Ser Leu Glu Ala Gln                                              Glu Tyr Arg Leu Tyr Arg Glu Lys                                                                 70                                                               75                   80                                                    - AAA TCA GCA TCT TGG ATT ACA                                               CGG ATA CGA CCA GAG CTT GTG AAG                                               AAC 464                                 Lys Ser Ala Ser Trp Ile Thr Arg Ile Arg Pro Glu Leu Val Lys Asn                                                                   85                                                               90                   95                 - GGC CAG TTC CAC ATC CCA TCC ATC ACC TGG GAA CAC ACA GGG CGA TAT 512                                               Gly Gln Phe His Ile Pro Ser Ile                                              Thr Trp Glu His Thr Gly Arg Tyr                                                        100                 105                                                             110                       - GGC TGT CAG TAT TAC AGC CGC GCT CGG TGG TCT GAG CTC AGT GAC CCC 560                                               Gly Cys Gln Tyr Tyr Ser Arg Ala                                              Arg Trp Ser Glu Leu Ser Asp Pro                                                    115                 120                                                             125                           - CTG GTG CTG GTG ATG ACA GGA GCC TAC CCA AAA CCC ACC CTC TCA GCC 608                                               Leu Val Leu Val Met Thr Gly Ala                                              Tyr Pro Lys Pro Thr Leu Ser Ala                                                130                 135                                                             140                 145                                                  - CAG CCC AGC CCT GTG GTG ACC                                               TCA GGA GGA AGG GTG ACC CTC CAG                                               TGT 656                                 Gln Pro Ser Pro Val Val Thr Ser Gly Gly Arg Val Thr Leu Gln Cys                                                                      150                                                             155                 160                                                      - GAG TCA CAG GTG GCA TTT GGC                                               GGC TTC ATT CTG TGT AAG GAA GGA                                               GAA 704                                 Glu Ser Gln Val Ala Phe Gly Gly Phe Ile Leu Cys Lys Glu Gly Glu                                                                  165                                                              170                 175                  - GAT GAA CAC CCA CAA TGC CTG AAC TCC CAG CCC CAT GCC CGT GGG TCG 752                                               Asp Glu His Pro Gln Cys Leu Asn                                              Ser Gln Pro His Ala Arg Gly Ser                                                        180                 185                                                             190                       - TCC CGC GCC ATC TTC TCC GTG GGC CCC GTG AGC CCG AAT CGC AGG TGG 800                                               Ser Arg Ala Ile Phe Ser Val Gly                                              Pro Val Ser Pro Asn Arg Arg Trp                                                    195                 200                                                            205                            - TCG CAC AGG TGC TAT GGT TAT GAC TTG AAC TCT CCC TAT GTG TGG TCT 848                                               Ser His Arg Cys Tyr Gly Tyr Asp                                              Leu Asn Ser Pro Tyr Val Trp Ser                                                210                 215                                                             220                 225                                                  - TCA CCC AGT GAT CTC CTG GAG                                               CTC CTG GTC CCA GGT GTT TCT AAG                                               AAG 896                                 Ser Pro Ser Asp Leu Leu Glu Leu Leu Val Pro Gly Val Ser Lys Lys                                                                     230                                                             235                 240                                                       - CCA TCA CTC TCA GTG CAG CCG                                               GGT CCT GTC GTG GCC CCT GGG GAA                                               AGC 944                                 Pro Ser Leu Ser Val Gln Pro Gly Pro Val Val Ala Pro Gly Glu Ser                                                                  245                                                              250                 255                  - CTG ACC CTC CAG TGT GTC TCT GAT GTC GGC TAT GAC AGA TTT GTT CTG 992                                               Leu Thr Leu Gln Cys Val Ser Asp                                              Val Gly Tyr Asp Arg Phe Val Leu                                                       260                 265                                                             270                        - TAC AAG GAG GGG GAA CGT GAC CTT CGC CAG CTC CCT GGC CGG CAG CCC 1040       Tyr Lys Glu Gly Glu Arg Asp Leu Arg Gln Leu Pro Gly Arg Gln Pro                                                          275                 280                                                             285                           - CAG GCT GGG CTC TCC CAG GCC AAC TTC ACC CTG GGC CCT GTG AGC CGC 1088       Gln Ala Gly Leu Ser Gln Ala Asn Phe Thr Leu Gly Pro Val Ser Arg                                                      290                 295                                                             300                 305                                                  - TCC TAC GGG GGC CAG TAC AGA                                               TGC TAC GGT GCA TAC AAC CTC TCC                                               TCC 1136                                Ser Tyr Gly Gly Gln Tyr Arg Cys Tyr Gly Ala Tyr Asn Leu Ser Ser                                                                      310                                                             315                 320                                                      - GAG TGG TCG GCC CCC AGC GAC                                               CCC CTG GAC ATC CTG ATC ACA GGA                                               CAG 1184                                Glu Trp Ser Ala Pro Ser Asp Pro Leu Asp Ile Leu Ile Thr Gly Gln                                                                  325                                                              330                 335                  - ATC CAT GGC ACA CCC TTC ATC TCA GTG CAG CCA GGC CCC ACA GTG GCC 1232       Ile His Gly Thr Pro Phe Ile Ser Val Gln Pro Gly Pro Thr Val Ala                                                              340                 345                                                             350                       - TCA GGA GAG AAC GTG ACC CTG CTG TGT CAG TCA TGG CGG CAG TTC CAC 1280       Ser Gly Glu Asn Val Thr Leu Leu Cys Gln Ser Trp Arg Gln Phe His                                                          355                 360                                                             365                           - ACT TTC CTT CTG ACC AAG GCG GGA GCA GCT GAT GCC CCA CTC CGT CTA 1328       Thr Phe Leu Leu Thr Lys Ala Gly Ala Ala Asp Ala Pro Leu Arg Leu                                                      370                 375                                                             380                 385                                                  - AGA TCA ATA CAC GAA TAT CCT                                               AAG TAC CAG GCT GAA TTC CCC ATG                                               AGT 1376                                Arg Ser Ile His Glu Tyr Pro Lys Tyr Gln Ala Glu Phe Pro Met Ser                                                                      390                                                             395                 400                                                      - CCT GTG ACC TCA GCC CAC GCG                                               GGG ACC TAC AGG ACC CTC CAT GGG                                               TTC 1424                                Pro Val Thr Ser Ala His Ala Gly Thr Tyr Arg Thr Leu His Gly Phe                                                                 405                                                               410                 415                  - CAG CCC CCC ACC CAC CGG TCC CAT CTC CAC ACC TGC AGG CCC 1466                                                      Gln Pro Pro Thr His Arg Ser His                                              Leu His Thr Cys Arg Pro                         420                 425                 430                            - TGAGGACCAG CCCCTCACCC CCACTGGGTC GGATCCCCAA AGTGGTCTGG GAAGGCACCT                                                1526                                     - GGGGGTTGTG ATCGGCATCT TGGTGGCCGT CGTCCTACTG CTCCTCCTCC TCCTCCTCCT                                                1586                                     - CTTCCTCATC CTCCGACATC GACGTCAGGG CAAACACTGG ACATCGACCC AGAGAAAGGC                                                1646                                     - TGATTTCCAA CATCCTGCAG GGGCTGTGGG GCCAGAGCCC ACAGACAGAG GCCTGCAGTG                                                1706                                     - GAGGTCCAGC CCAGCTGCCG ACGCCCAGGA AGAAAACCTC TATGCTGCCG TGAAGGACAC                                                1766                                     - ACAGCCTGAA GATGGGGTGG AGATGGACAC TCGGGCTGCT GCATCTGAAG CCCCCCAGGA                                                1826                                     - TGTGACCTAC GCCCAGCTGC ACAGCTTGAC CCTCAGACGG AAGGCAACTG AGCCTCCTCC                                                1886                                     - ATCCCAGGAA AGGGAACCTC CAGCTGAGCC CAGCATCTAC GCCACCCTGG CCATCCACTA                                                1946                                     - GCCCGGAGGG TACGCAGACT CCACACTCAG TAGAAGGAGA CTCAGGACTG CTGAAGGCAC                                                2006                                     - GGGAGCTGCC CCCAGTGGAC ACCAATGAAC CCCAGTCAGC CTGGACCCCT AACAAAGACC                                                2066                                     - ATGAGGAGAT GCTGGGAACT TTGGGACTCA CTTGATTCTG CAGTCGAAAT AACTAATATC                                                2126                                     - CCTACATTTT TTAATTAAAG CAACAGACTT CTCAATAATC AATGAGTTAA CCGAGAAAAC                                                2186                                     - TAAAAAAAAA AAAA 2200                                                        -                                                                          A KTE03 embodiment designated KLM67 (SEQ ID NO: 21 and 22):                   GCCACACGCA GCTCAGCCTG GGCGGCACAG CCAGATGCGA GATGCGTCTC TGCTGATCTG                                                     60                                       - AGTCTGCCTG CAGCATGGAC CTGGGTCTTC CCTGAAGCAT CTCCAGGGCT GGAGGGACGA                                                120                                      - CTGCCATGCA CCGAGGGCTC ATCCATCCAC AGAGCAGGGC AGTGGGAGGA GACGCC 176                                                  - ATG ACC CCC ATC CTC ACG GTC                                               CTG ATC TGT CTC GGG CTG AGT CTG                                               GGC 224                                 Met Thr Pro Ile Leu Thr Val Leu Ile Cys Leu Gly Leu Ser Leu Gly                                                        1              5                                                              10                  15                                                       - CCC CGG ACC CAC GTG CAG GCA                                               GGG CAC CTC CCC AAG CCC ACC CTC                                               TGG 272                                 Pro Arg Thr His Val Gln Ala Gly His Leu Pro Lys Pro Thr Leu Trp                                                                   20                                                              25                  30                   - GCT GAA CCA GGC TCT GTG ATC ACC CAG GGG AGT CCT GTG ACC CTC AGG 320                                               Ala Glu Pro Gly Ser Val Ile Thr                                              Gln Gly Ser Pro Val Thr Leu Arg                                                         35                  40                                                              45                       - TGT CAG GGG GGC CAG GAG ACC CAG GAG TAC CGT CTA TAT AGA GAA AAG 368                                               Cys Gln Gly Gly Gln Glu Thr Gln                                              Glu Tyr Arg Leu Tyr Arg Glu Lys                                                     50                  55                                                              60                           - AAA ACA GCA CCC TGG ATT ACA CGG ATC CCA CAG GAG CTT GTG AAG AAG 416                                               Lys Thr Ala Pro Trp Ile Thr Arg                                              Ile Pro Gln Glu Leu Val Lys Lys                                                 65                  70                                                              75                  80                                                  - GGC CAG TTC CCC ATC CCA TCC                                               ATC ACC TGG GAA CAT GCA GGG CGG                                               TAT 464                                 Gly Gln Phe Pro Ile Pro Ser Ile Thr Trp Glu His Ala Gly Arg Tyr                                                                       85                                                              90                  95                                                      - CGC TGT TAC TAT GGT AGC GAC                                               ACT GCA GGC CGC TCA GAG AGC AGT                                               GAC 512                                 Arg Cys Tyr Tyr Gly Ser Asp Thr Ala Gly Arg Ser Glu Ser Ser Asp                                                                  100                                                              105                 110                  - CCC CTG GAG CTG GTG GTG ACA GGA GCC TAC ATC AAA CCC ACC CTC TCA 560                                               Pro Leu Glu Leu Val Val Thr Gly                                              Ala Tyr Ile Lys Pro Thr Leu Ser                                                        115                 120                                                             125                       - GCC CAG CCC AGC CCC GTG GTG AAC TCA GGA GGG AAT GTA ACC CTC CAG 608                                               Ala Gln Pro Ser Pro Val Val Asn                                              Ser Gly Gly Asn Val Thr Leu Gln                                                    130                 135                                                             140                           - TGT GAC TCA CAG GTG GCA TTT GAT GGC TTC ATT CTG TGT AAG GAA GGA 656                                               Cys Asp Ser Gln Val Ala Phe Asp                                              Gly Phe Ile Leu Cys Lys Glu Gly                                                145                 150                                                             155                 160                                                  - GAA GAT GAA CAC CCA CAA TGC                                               CTG AAC TCC CAG CCC CAT GCC CGT                                               GGG 704                                 Glu Asp Glu His Pro Gln Cys Leu Asn Ser Gln Pro His Ala Arg Gly                                                                      165                                                             170                 175                                                      - TCG TCC CGC GCC ATC TTC TCC                                               GTG GGC CCC GTG AGC CCG AGT CGC                                               AGG 752                                 Ser Ser Arg Ala Ile Phe Ser Val Gly Pro Val Ser Pro Ser Arg Arg                                                                  180                                                              185                 190                  - TGG TGG TAC AGG TGC TAT GCT TAT GAC TCG AAC TCT CCC TAT GAG TGG 800                                               Trp Trp Tyr Arg Cys Tyr Ala Tyr                                              Asp Ser Asn Ser Pro Tyr Glu Trp                                                        195                 200                                                             205                       - TCT CTA CCC AGT GAT CTC CTG GAG CTC CTG GTC CTA GGT GTT TCT AAG 848                                               Ser Leu Pro Ser Asp Leu Leu Glu                                              Leu Leu Val Leu Gly Val Ser Lys                                                    210                 215                                                             220                           - AAG CCA TCA CTC TCA GTG CAG CCA GGT CCT ATC GTG GCC CCT GAG GAG 896                                               Lys Pro Ser Leu Ser Val Gln Pro                                              Gly Pro Ile Val Ala Pro Glu Glu                                                225                 230                                                             235                 240                                                  - ACC CTG ACT CTG CAG TGT GGC                                               TCT GAT GCT GGC TAC AAC AGA TTT                                               GTT 944                                 Thr Leu Thr Leu Gln Cys Gly Ser Asp Ala Gly Tyr Asn Arg Phe Val                                                                      245                                                             250                 255                                                      - CTG TAT AAG GAC GGG GAA CGT                                               GAC TTC CTT CAG CTC GCT GGC GCA                                               CAG 992                                 Leu Tyr Lys Asp Gly Glu Arg Asp Phe Leu Gln Leu Ala Gly Ala Gln                                                                  260                                                              265                 270                  - CCC CAG GCT GGG CTC TCC CAG GCC AAC TTC ACC CTG GGC CCT GTG AGC 1040       Pro Gln Ala Gly Leu Ser Gln Ala Asn Phe Thr Leu Gly Pro Val Ser                                                              275                 280                                                             285                       - CGC TCC TAC GGG GGC CAG TAC AGA TGC TAC GGT GCA CAC AAC CTC TCC 1088       Arg Ser Tyr Gly Gly Gln Tyr Arg Cys Tyr Gly Ala His Asn Leu Ser                                                          290                 295                                                             300                           - TCC GAG TGG TCG GCC CCC AGC GAC CCC CTG GAC ATC CTG ATC GCA GGA 1136       Ser Glu Trp Ser Ala Pro Ser Asp Pro Leu Asp Ile Leu Ile Ala Gly                                                      305                 310                                                             315                 320                                                  - CAG TTC TAT GAC AGA GTC TCC                                               CTC TCG GTG CAG CCG GGC CCC ACG                                               GTG 1184                                Gln Phe Tyr Asp Arg Val Ser Leu Ser Val Gln Pro Gly Pro Thr Val                                                                      325                                                             330                 335                                                      - GCC TCA GGA GAG AAC GTG ACC                                               CTG CTG TGT CAG TCA CAG GGA TGG                                               ATG 1232                                Ala Ser Gly Glu Asn Val Thr Leu Leu Cys Gln Ser Gln Gly Trp Met                                                                  340                                                              345                 350                  - CAA ACT TTC CTT CTG ACC AAG GAG GGG GCA GCT GAT GAC CCA TGG CGT 1280       Gln Thr Phe Leu Leu Thr Lys Glu Gly Ala Ala Asp Asp Pro Trp Arg                                                              355                 360                                                             365                       - CTA AGA TCA ACG TAC CAA TCT CAA AAA TAC CAG GCT GAA TTC CCC ATG 1328       Leu Arg Ser Thr Tyr Gln Ser Gln Lys Tyr Gln Ala Glu Phe Pro Met                                                          370                 375                                                             380                           - GGT CCT GTG ACC TCA GCC CAT GCG GGG ACC TAC AGG TGC TAC GGC TCA 1376       Gly Pro Val Thr Ser Ala His Ala Gly Thr Tyr Arg Cys Tyr Gly Ser                                                      385                 390                                                             395                 400                                                  - CAG AGC TCC AAA CCC TAC CTG                                               CTG ACT CAC CCC AGT GAC CCC CTG                                               GAG 1424                                Gln Ser Ser Lys Pro Tyr Leu Leu Thr His Pro Ser Asp Pro Leu Glu                                                                      405                                                             410                 415                                                      - CTC GTG GTC TCA GGA CCG TCT                                               GGG GGC CCC AGC TCC CCG ACA ACA                                               GGC 1472                                Leu Val Val Ser Gly Pro Ser Gly Gly Pro Ser Ser Pro Thr Thr Gly                                                                  420                                                              425                 430                  - CCC ACC TCC ACA TCT GGC CCT GAG GAC CAG CCC CTC ACC CCC ACC GGG 1520       Pro Thr Ser Thr Ser Gly Pro Glu Asp Gln Pro Leu Thr Pro Thr Gly                                                              435                 440                                                             445                       - TCG GAT CCC CAG AGT GGT CTG GGA AGG CAC CTG GGG GTT GTG ATC GGC 1568       Ser Asp Pro Gln Ser Gly Leu Gly Arg His Leu Gly Val Val Ile Gly                                                          450                 455                                                             460                           - ATC TTG GTG GCC GTC ATC CTA CTG CTC CTC CTC CTC CTC CTC CTC TTC 1616       Ile Leu Val Ala Val Ile Leu Leu Leu Leu Leu Leu Leu Leu Leu Phe                                                      465                 470                                                             475                 480                                                  - CTC ATC CTC CGA CAT CGA CGT                                               CAG GGC AAA CAC TGG ACA TCG ACC                                               CAG 1664                                Leu Ile Leu Arg His Arg Arg Gln Gly Lys His Trp Thr Ser Thr Gln                                                                      485                                                             490                 495                                                      - AGA AAG GCT GAT TTC CAA CAT                                               CCT GCA GGG GCT GTG GGG CCA GAG                                               CCC 1712                                Arg Lys Ala Asp Phe Gln His Pro Ala Gly Ala Val Gly Pro Glu Pro                                                                  500                                                              505                 510                  - ACA GAC AGA CGC CTG CAG TGG AGG TCC AGC CCA GCT GCC GAT GCC CAG 1760       Thr Asp Arg Arg Leu Gln Trp Arg Ser Ser Pro Ala Ala Asp Ala Gln                                                              515                 520                                                             525                       - GAA GAA AAC CTC TAT GCT GCC GTG AAG CAC ACA CAG CCT GAG GAT GGG 1808       Glu Glu Asn Leu Tyr Ala Ala Val Lys His Thr Gln Pro Glu Asp Gly                                                          530                 535                                                             540                           - GTG GAG ATG GAC ACT CGG CAG AGC CCA CAC GAT GAA GAC CCC CAG GCA 1856       Val Glu Met Asp Thr Arg Gln Ser Pro His Asp Glu Asp Pro Gln Ala                                                      545                 550                                                             555                 560                                                  - GTG ACG TAT GCC GAG GTG AAA                                               CAC TCC AGA CCT AGG AGA GAA ATG                                               GCT 1904                                Val Thr Tyr Ala Glu Val Lys His Ser Arg Pro Arg Arg Glu Met Ala                                                                      565                                                             570                 575                                                      - TCT CCT CCT TCC CCA CTG TCT                                               GGG GAA TTC CTG GAC ACA AAG GAC                                               AGA 1952                                Ser Pro Pro Ser Pro Leu Ser Gly Glu Phe Leu Asp Thr Lys Asp Arg                                                                  580                                                              585                 590                  - CAG GCG GAA GAG GAC AGG CAG ATG GAC ACT GAG GCT GCT GCA TCT GAA 2000       Gln Ala Glu Glu Asp Arg Gln Met Asp Thr Glu Ala Ala Ala Ser Glu                                                              595                 600                                                             605                       - GCC CCC CAG GAT GTG ACC TAC GCC CAG CTG CAC AGC TTG ACC CTT AGA 2048       Ala Pro Gln Asp Val Thr Tyr Ala Gln Leu His Ser Leu Thr Leu Arg                                                          610                 615                                                             620                           - CGG AAG GCA ACT GAG CCT CCT CCA TCC CAG GAA GGG CCC TCT CCA GCT 2096       Arg Lys Ala Thr Glu Pro Pro Pro Ser Gln Glu Gly Pro Ser Pro Ala                                                      625                 630                                                             635                 640                                                  - GTG CCC AGC ATC TAC GCC ACT                                               CTG GCC ATC CAC TAG CCCAGGGGGG                                                2142                                    Val Peo Ser Ile Tyr Ala Thr Leu Ala Ile His  *                                                645                 650                                        - GACGCAGACC CCACACTCCA TGGAGTCTGG AATGCATGGG AGCTGCCCCC CCAGTGGACA                                                2202                                     - CCATTGGACC CCACCCAGCC TGGATCTACC CCAGGAGACT CTGGGAACTT TTAGGGGTCA                                                2262                                     - CTCAATTCTG CAGTATAAAT AACTAATGTC TCTACAATTT TGAAATAAAG CAACAGACTT                                                2322                                     - CTCAATAATC AATGAAGTAG CTGAGAAAAC TAAGTCAGAA AGTGCATTAA ACTGAATCAC                                                2382                                     - AATGTAAATA TTACACATCA AGCGATGAAA CTGGAAAACT ACAAGCCACG AATGAATGAA                                                2442                                     - TTAGGAAAGA AAAAAAGTAG GAAATGAATG ATCTTGGCTT TCCTATAAGA AATTTAGGGC                                                2502                                     - AGGGCACGGT CGCTCACGCC TGTAATTCCA GCACTTTGGG AGGCCGAGGC GGGCAGATCA                                                2562                                     - CGAGTTCAGG AGATCGAGAC CATCTTGGCC AACATGGTGA AACCCTGTCT CTCCTAAAAA                                                2622                                     - TACAAAAATT AGCTGGATGT GGTGGCAGTG CCTGTAATCC CAGCTATTTG GGAGGCTGAG                                                2682                                     - GCAGGAGAAT CGCTTGAACC AGGGAGTCAG AGGTTTCAGT GAGCCAAGAT CGCACCACTG                                                2742                                     - CTCTCCAGCC TGGCGACAGA GGGAGACTCC ATCTCAAATT AAAAAAAA 2790                __________________________________________________________________________

The peptide segments can also be used to produce appropriateoligonucleotides to screen a library to determine the presence of asimilar gene, e.g., an identical or polymorphic variant, or to identifya monocyte. The genetic code can be used to select appropriateoligonucleotides useful as probes for screening. In combination withpolymerase chain reaction (PCR) techniques, synthetic oligonucleotideswill be useful in selecting desired clones from a library.

Complementary sequences will also be used as probes or primers. Basedupon identification of the likely amino terminus, other peptides shouldbe particularly useful, e.g., coupled with anchored vector or poly-Acomplementary PCR techniques or with complementary DNA of otherpeptides.

Techniques for nucleic acid manipulation of genes encoding thesemonocyte proteins, e.g., subcloning nucleic acid sequences encodingpolypeptides into expression vectors, labeling probes, DNAhybridization, and the like are described generally in Sambrook, et al.(1989) Molecular Cloning: A Laboratory Manual (2nd ed.) Vol. 1-3, ColdSpring Harbor Laboratory, Cold Spring Harbor Press, NY, which isincorporated herein by reference and hereinafter referred to as"Sambrook, et al." See also, Coligan, et al. (1987 and periodicsupplements) Current Protocols in Molecular Biology Greene/Wiley, NewYork, N.Y., referred to as "Coligan, et al."

There are various methods of isolating the DNA sequences encoding thesemonocyte proteins. For example, DNA is isolated from a genomic or cDNAlibrary using labeled oligonucleotide probes having sequences identicalor complementary to the sequences disclosed herein. Full-length probesmay be used, or oligonucleotide probes may be generated by comparison ofthe sequences disclosed with other proteins and selecting specificprimers. Such probes can be used directly in hybridization assays toisolate DNA encoding monocyte proteins, or probes can be designed foruse in amplification techniques such as PCR, for the isolation of DNAencoding monocyte proteins.

To prepare a cDNA library, mRNA is isolated from cells which express themonocyte protein. cDNA is prepared from the mRNA and ligated into arecombinant vector. The vector is transfected into a recombinant hostfor propagation, screening and cloning. Methods for making and screeningcDNA libraries are well known. See Gubler and Hoffman (1983) Gene25:263-269; Sambrook, et al.; or Coligan, et al.

For a genomic library, the DNA can be extracted from tissue and eithermechanically sheared or enzymatically digested to yield fragments ofabout 12-20 kb. The fragments are then separated by gradientcentrifugation and cloned in bacteriophage lambda vectors. These vectorsand phage are packaged in vitro, as described, e.g., in Sambrook, et al.or Coligan, et al. Recombinant phage are analyzed by plaquehybridization as described in Benton and Davis (1977) Science196:180-182. Colony hybridization is carried out as generally describedin, e.g., Grunstein, et al. (1975) Proc. Natl. Acad. Sci. USA72:3961-3965.

DNA encoding a monocyte protein can be identified in either cDNA orgenomic libraries by its ability to hybridize with the nucleic acidprobes described herein, for example in colony or plaque hybridizationexperiments. The corresponding DNA regions are isolated by standardmethods familiar to those of skill in the art. See Sambrook, et al.

Various methods of amplifying target sequences, such as the polymerasechain reaction, can also be used to prepare DNA encoding monocyteproteins. Polymerase chain reaction (PCR) technology is used to amplifysuch nucleic acid sequences directly from mRNA, from cDNA, and fromgenomic libraries or cDNA libraries. The isolated sequences encodingmonocyte proteins may also be used as templates for PCR amplification.

In PCR techniques, oligonucleotide primers complementary to two 5'regions in the DNA region to be amplified are synthesized. Thepolymerase chain reaction is then carried out using the two primers. SeeInnis, et al. (eds.) (1990) PCR Protocols: A Guide to Methods andApplications Academic Press, San Diego, Calif. Primers can be selectedto amplify the entire regions encoding a selected full-length monocyteprotein or to amplify smaller DNA segments as desired. Once such regionsare PCR-amplified, they can be sequenced and oligonucleotide probes canbe prepared from sequence obtained using standard techniques. Theseprobes can then be used to isolate DNAs encoding other forms of themonocyte proteins.

Oligonucleotides for use as probes are chemically synthesized accordingto the solid phase phosphoramidite triester method first described byBeaucage and Carruthers (1983) Tetrahedron Lett. 22(20):1859-1862, orusing an automated synthesizer, as described in Needham-VanDevanter, etal. (1984) Nucleic Acids Res. 12:6159-6168. Purification ofoligonucleotides is performed e.g., by native acrylamide gelelectrophoresis or by anion-exchange HPLC as described in Pearson andRegnier (1983) J. Chrom. 255:137-149. The sequence of the syntheticoligonucleotide can be verified using the chemical degradation method ofMaxam and Gilbert in Grossman and Moldave (eds.) (1980) Methods inEnzymology 65:499-560 Academic Press, New York.

An isolated nucleic acid encoding a human protein which is a type Itransmembrane protein comprising an extracellular portion characterizedby Ig-like domains, indicating that this gene encodes a receptor memberof the Ig superfamily. This clone has been designated FDF03. Itsnucleotide sequence and corresponding open reading frame are provided inSEQ ID NO: 1 and 2, respectively. An N-terminal hydrophobic sequence,e.g., a putative signal sequence, corresponds to about amino acidresidues -19(met) to -1(leu), and a internal hydrophobic segment,corresponding to a putative transmembrane segment runs from aroundala177 to leu199. Other mammalian counterparts should become available,e.g., a partial rodent gene is described in SEQ ID NO: 3 and 4. Standardtechniques will allow isolation of other counterparts, or to extendpartial sequences.

A second human monocyte cell clone was isolated, designated YE01, isrelated to the receptors for Fc gamma and/or Fc alpha. This has alsobeen referred to as DNAX Leukocyte Associated Immunoglobulin-likeReceptor (DLAIR). See also Meyaard, et al. (1997) Immunity 7:283-290,which was published by the inventors after the priority date of thisapplication, and is incorporated herein by reference. This protein isreferred to herein as an Fc gamma/alpha receptor and is described in SEQID NO: 5 and 6. Another human isolate is described in SEQ ID NO: 7 and8. A soluble form of the receptor is encoded in SEQ ID NO: 9 and 10.While the gene was initially described as a monocyte derived gene,expression analysis indicates that it is more specific for expression onlymphocytes. Thus, in the case of YE01, the descriptor "monocyte gene"may indicate its original identification in a population enriched forthat cell type, though it may have also contained some other cell types.Sequence analysis suggests YE01 is a member of the Ig superfamily ofreceptors, and is closely related to the CD8 family, which contain aV1J-type fold, particularly the Fc receptors alpha and/or gamma. Becauseit contains an ITIM (immune receptor tyrosine-based inhibitory motif)like motif, the protein may well be a lymphocyte version of the KillerInhibitory Receptors (KIR), which send a negative signal to inhibitkiller cell function. This protein exhibits similar function ininhibiting lymphocyte effector function, e.g., antigen presentation orsubsequent response initiation.

In particular, signaling through the molecule recognized by DX26 mAb(designated DNAX Leukocyte Associated Immunoglobulin-like Receptor(DLAIR)), delivers a negative signal to NK cell clones that preventstheir killing specific target cells. However, the molecule is expressedon other lymphocytes, including T cells and monocytes. Thus, the DX26antibody probably represents an antibody which both inhibits NK andcytotoxic T cell killing, and the monocyte distribution suggests thatthe molecule may inhibit monocyte-mediated or lymphocyte-mediatedeffector functions.

A third monocyte gene was isolated and designated KTE03, and isrepresented by six related embodiments, designated YYB01, YYB04 (forms 1and 2), (KIR-Like Molecule) KLM63, KLM66, and KLM67. See SEQ ID NO:11-22. Note that a possible splice variant, which may encode a variantprotein form, has been detected.

This invention provides isolated DNA or fragments to encode a monocyteprotein, as described. In addition, this invention provides isolated orrecombinant DNA which encodes a biologically active protein orpolypeptide which is capable of hybridizing under appropriateconditions, e.g., high stringency, with the DNA sequences describedherein. Said biologically active protein or polypeptide can be anaturally occurring form, or a recombinant protein or fragment, and havean amino acid sequence as disclosed in SEQ ID NO: 2 or 4; 6, 8, or 10;or 12, 14, 16, 18, 20, or 22. Preferred embodiments will be full lengthnatural isolates, e.g., from a primate. In glycosylated form, theproteins should exhibit larger sizes. Further, this inventionencompasses the use of isolated or recombinant DNA, or fragmentsthereof, which encode proteins which are homologous to each respectivemonocyte protein. The isolated DNA can have the respective regulatorysequences in the 5' and 3' flanks, e.g., promoters, enhancers, poly-Aaddition signals, and others.

IV. Making Monocyte Gene Products

DNAs which encode these monocyte proteins or fragments thereof can beobtained by chemical synthesis, screening cDNA libraries, or byscreening genomic libraries prepared from a wide variety of cell linesor tissue samples.

These DNAs can be expressed in a wide variety of host cells for thesynthesis of a full-length protein or fragments which can, e.g., be usedto generate polyclonal or monoclonal antibodies; for binding studies;for construction and expression of modified molecules; and forstructure/function studies. Each of these monocyte proteins or theirfragments can be expressed in host cells that are transformed ortransfected with appropriate expression vectors. These molecules can besubstantially purified to be free of protein or cellular contaminants,other than those derived from the recombinant host, and therefore areparticularly useful in pharmaceutical compositions when combined with apharmaceutically acceptable carrier and/or diluent. The antigen, orportions thereof, may be expressed as fusions with other proteins.

Expression vectors are typically self-replicating DNA or RNA constructscontaining the desired monocyte gene or its fragments, usually operablylinked to suitable genetic control elements that are recognized in asuitable host cell. These control elements are capable of effectingexpression within a suitable host. The specific type of control elementsnecessary to effect expression will depend upon the eventual host cellused. Generally, the genetic control elements can include a prokaryoticpromoter system or a eukaryotic promoter expression control system, andtypically include a transcriptional promoter, an optional operator tocontrol the onset of transcription, transcription enhancers to elevatethe level of mRNA expression, a sequence that encodes a suitableribosome binding site, and sequences that terminate transcription andtranslation. Expression vectors also usually contain an origin ofreplication that allows the vector to replicate independently from thehost cell.

The vectors of this invention contain DNAs which encode the variousmonocyte proteins, or a fragment thereof, typically encoding, e.g., abiologically active polypeptide, or protein. The DNA can be under thecontrol of a viral promoter and can encode a selection marker. Thisinvention further contemplates use of such expression vectors which arecapable of expressing eukaryotic cDNA coding for a monocyte protein in aprokaryotic or eukaryotic host, where the vector is compatible with thehost and where the eukaryotic cDNA coding for the protein is insertedinto the vector such that growth of the host containing the vectorexpresses the cDNA in question. Usually, expression vectors are designedfor stable replication in their host cells or for amplification togreatly increase the total number of copies of the desirable gene percell. It is not always necessary to require that an expression vectorreplicate in a host cell, e.g., it is possible to effect transientexpression of the protein or its fragments in various hosts usingvectors that do not contain a replication origin that is recognized bythe host cell. It is also possible to use vectors that cause integrationof a monocyte gene or its fragments into the host DNA by recombination,or to integrate a promoter which controls expression of an endogenousgene.

Vectors, as used herein, comprise plasmids, viruses, bacteriophage,integratable DNA fragments, and other vehicles which enable theintegration of DNA fragments into the genome of the host. Expressionvectors are specialized vectors which contain genetic control elementsthat effect expression of operably linked genes. Plasmids are the mostcommonly used form of vector but all other forms of vectors which servean equivalent function are suitable for use herein. See, e.g., Pouwels,et al. (1985 and Supplements) Cloning Vectors: A Laboratory ManualElsevier, N.Y.; and Rodriquez, et al. (eds.) (1988) Vectors: A Survey ofMolecular Cloning Vectors and Their Uses Buttersworth, Boston, Mass.

Suitable host cells include prokaryotes, lower eukaryotes, and highereukaryotes. Prokaryotes include both gram negative and gram positiveorganisms, e.g., E. coli and B. subtilis. Lower eukaryotes includeyeasts, e.g., S. cerevisiae and Pichia, and species of the genusDictyostelium. Higher eukaryotes include established tissue culture celllines from animal cells, both of non-mammalian origin, e.g., insectcells, and birds, and of mammalian origin, e.g., human, primates, androdents.

Prokaryotic host-vector systems include a wide variety of vectors formany different species. As used herein, E. coli and its vectors will beused generically to include equivalent vectors used in otherprokaryotes. A representative vector for amplifying DNA is pBR322 or itsderivatives. Vectors that can be used to express monocyte proteins orfragments include, but are not limited to, such vectors as thosecontaining the lac promoter (pUC-series); trp promoter (pBR322-trp); Ipppromoter (the pIN-series); lambda-pP or pR promoters (pOTS); or hybridpromoters such as ptac (pDR540). See Brosius, et al. (1988) "ExpressionVectors Employing Lambda-, trp-, lac-, and Ipp-derived Promoters", inRodriguez and Denhardt (eds.) Vectors: A Survey of Molecular CloningVectors and Their Uses 10:205-236 Buttersworth, Boston, Mass.

Lower eukaryotes, e.g., yeasts and Dictyostelium, may be transformedwith monocyte gene sequence containing vectors. For purposes of thisinvention, the most common lower eukaryotic host is the baker's yeast,Saccharomyces cerevisiae. It will be used generically to represent lowereukaryotes although a number of other strains and species are alsoavailable. Yeast vectors typically consist of a replication origin(unless of the integrating type), a selection gene, a promoter, DNAencoding the desired protein or its fragments, and sequences fortranslation termination, polyadenylation, and transcription termination.Suitable expression vectors for yeast include such constitutivepromoters as 3-phosphoglycerate kinase and various other glycolyticenzyme gene promoters or such inducible promoters as the alcoholdehydrogenase 2 promoter or metallothionine promoter. Suitable vectorsinclude derivatives of the following types: self-replicating low copynumber (such as the YRp-series), self-replicating high copy number (suchas the YEp-series); integrating types (such as the YIp-series), ormini-chromosomes (such as the YCp-series).

Higher eukaryotic tissue culture cells are the preferred host cells forexpression of the monocyte protein. In principle, most any highereukaryotic tissue culture cell line may be used, e.g., insectbaculovirus expression systems, whether from an invertebrate orvertebrate source. However, mammalian cells are preferred to achieveproper processing, both cotranslationally and posttranslationally.Transformation or transfection and propagation of such cells is routine.Useful cell lines include HeLa cells, Chinese hamster ovary (CHO) celllines, baby rat kidney (BRK) cell lines, insect cell lines, bird celllines, and monkey (COS) cell lines. Expression vectors for such celllines usually include an origin of replication, a promoter, atranslation initiation site, RNA splice sites (e.g., if genomic DNA isused), a polyadenylation site, and a transcription termination site.These vectors also may contain a selection gene or amplification gene.Suitable expression vectors may be plasmids, viruses, or retrovirusescarrying promoters derived, e.g., from such sources as from adenovirus,SV40, parvoviruses, vaccinia virus, or cytomegalovirus. Representativeexamples of suitable expression vectors include pcDNA1; pCD, seeOkayama, et al. (1985) Mol. Cell Biol. 5:1136-1142; pMClneo Poly-A, seeThomas, et al. (1987) Cell 51:503-512; and a baculovirus vector such aspAC 373 or pAC 610.

In certain instances, the monocyte proteins need not be glycosylated toelicit biological responses in certain assays. However, it will often bedesirable to express a monocyte polypeptide in a system which provides aspecific or defined glycosylation pattern. In this case, the usualpattern will be that provided naturally by the expression system.However, the pattern will be modifiable by exposing the polypeptide,e.g., in unglycosylated form, to appropriate glycosylating proteinsintroduced into a heterologous expression system. For example, amonocyte gene may be co-transformed with one or more genes encodingmammalian or other glycosylating enzymes. It is further understood thatover glycosylation may be detrimental to monocyte protein biologicalactivity, and that one of skill may perform routine testing to optimizethe degree of glycosylation which confers optimal biological activity.

A monocyte protein, or a fragment thereof, may be engineered to bephosphatidyl inositol (PI) linked to a cell membrane, but can be removedfrom membranes by treatment with a phosphatidyl inositol cleavingenzyme, e.g., phosphatidyl inositol phospholipase-C. This releases theantigen in a biologically active form, and allows purification bystandard procedures of protein chemistry. See, e.g., Low (1989) Biochem.Biophys. Acta 988:427-454; Tse, et al. (1985) Science 230:1003-1008;Brunner, et al. (1991) J. Cell Biol. 114:1275-1283; and Coligan, et al.(eds.) (1996 and periodic supplements) Current Protocols in ProteinScience, John Wiley & Sons, New York, N.Y.

Now that these monocyte proteins have been characterized, fragments orderivatives thereof can be prepared by conventional processes forsynthesizing peptides. These include processes such as are described inStewart and Young (1984) Solid Phase Peptide Synthesis Pierce ChemicalCo., Rockford, Ill.; Bodanszky and Bodanszky (1984) The Practice ofPeptide Synthesis Springer-Verlag, New York, N.Y.; and Bodanszky (1984)The Principles of Peptide Synthesis Springer-Verlag, New York, N.Y. Seealso Merrifield (1986) Science 232:341-347; and Dawson, et al. (1994)Science 266:776-779. For example, an azide process, an acid chlorideprocess, an acid anhydride process, a mixed anhydride process, an activeester process (for example, p-nitrophenyl ester, N-hydroxysuccinimideester, or cyanomethyl ester), a carbodiimidazole process, anoxidative-reductive process, or a dicyclohexylcarbodiimide(DCCD)/additive process can be used. Solid phase and solution phasesyntheses are both applicable to the foregoing processes.

The prepared protein and fragments thereof can be isolated and purifiedfrom the reaction mixture by means of peptide separation, for example,by extraction, precipitation, electrophoresis and various forms ofchromatography, and the like. The monocyte proteins of this inventioncan be obtained in varying degrees of purity depending upon the desireduse. Purification can be accomplished by use of known proteinpurification techniques or by the use of the antibodies or bindingpartners herein described, e.g., in immunoabsorbant affinitychromatography. This immunoabsorbant affinity chromatography is carriedout by first linking the antibodies to a solid support and contactingthe linked antibodies with solubilized lysates of appropriate sourcecells, lysates of other cells expressing the protein, or lysates orsupernatants of cells producing the proteins as a result of DNAtechniques, see below.

Multiple cell lines may be screened for one which expresses said proteinat a high level compared with other cells. Various cell lines, e.g., amouse thymic stromal cell line TA4, is screened and selected for itsfavorable handling properties. Natural monocyte cell proteins can beisolated from natural sources, or by expression from a transformed cellusing an appropriate expression vector. Purification of the expressedprotein is achieved by standard procedures, or may be combined withengineered means for effective purification at high efficiency from celllysates or supernatants. FLAG or His₆ segments can be used for suchpurification features.

V. Antibodies

Antibodies can be raised to these various monocyte proteins, includingindividual, polymorphic, allelic, strain, or species variants, andfragments thereof, both in their naturally occurring (full-length) formsand in their recombinant forms. Additionally, antibodies can be raisedto monocyte proteins in either their active forms or in their inactiveforms. Anti-idiotypic antibodies may also be used.

a. Antibody Production

A number of immunogens may be used to produce antibodies specificallyreactive with these monocyte proteins. Recombinant protein is thepreferred immunogen for the production of monoclonal or polyclonalantibodies. Naturally occurring protein may also be used either in pureor impure form. Synthetic peptides made using the human monocyte proteinsequences described herein may also used as an immunogen for theproduction of antibodies to the monocyte protein. Recombinant proteincan be expressed in eukaryotic or prokaryotic cells as described herein,and purified as described. The product is then injected into an animalcapable of producing antibodies. Either monoclonal or polyclonalantibodies may be generated for subsequent use in immunoassays tomeasure the protein.

Methods of producing polyclonal antibodies are known to those of skillin the art. In brief, an immunogen, preferably a purified protein, ismixed with an adjuvant and animals are immunized with the mixture. Theanimal's immune response to the immunogen preparation is monitored bytaking test bleeds and determining the titer of reactivity to themonocyte protein of interest. When appropriately high titers of antibodyto the immunogen are obtained, blood is collected from the animal andantisera are prepared. Further fractionation of the antisera to enrichfor antibodies reactive to the protein can be done if desired. See,e.g., Harlow and Lane.

Monoclonal antibodies may be obtained by various techniques familiar tothose skilled in the art. Briefly, spleen cells from an animal immunizedwith a desired antigen are immortalized, commonly by fusion with amyeloma cell. See, e.g., Kohler and Milstein (1976) Eur. J. Immunol.6:511-519, which is incorporated herein by reference. Alternativemethods of immortalization include transformation with Epstein BarrVirus, oncogenes, or retroviruses, or other methods known in the art.Colonies arising from single immortalized cells are screened forproduction of antibodies of the desired specificity and affinity for theantigen, and yield of the monoclonal antibodies produced by such cellsmay be enhanced by various techniques, including injection into theperitoneal cavity of a vertebrate host. Alternatively, one may isolateDNA sequences which encode a monoclonal antibody or a binding fragmentthereof by screening a DNA library from human B cells according to thegeneral protocol outlined by Huse, et al. (1989) Science 246:1275-1281.

Antibodies, including binding fragments and single chain versions,against predetermined fragments of these monocyte proteins can be raisedby immunization of animals with conjugates of the fragments with carrierproteins as described above. Monoclonal antibodies are prepared fromcells secreting the desired antibody. These antibodies can be screenedfor binding to normal or defective monocyte proteins, or screened foragonistic or antagonistic activity. These monoclonal antibodies willusually bind with at least a K_(D) of about 1 mM, more usually at leastabout 300 μM, typically at least about 100 μM, more typically at leastabout 30 μM, preferably at least about 10 μM, and more preferably atleast about 3 μM or better. Standard methods are available for selectionof high affinity and selective antibody preparations.

In some instances, it is desirable to prepare monoclonal antibodies fromvarious mammalian hosts, such as mice, rodents, primates, humans, etc.Description of techniques for preparing such monoclonal antibodies maybe found in, e.g., Stites, et al. (eds.) Basic and Clinical Immunology(4th ed.) Lange Medical Publications, Los Altos, Calif., and referencescited therein; Harlow and Lane (1988) Antibodies: A Laboratory ManualCSH Press; Goding (1986) Monoclonal Antibodies: Principles and Practice(2d ed.) Academic Press, New York, N.Y.; and particularly in Kohler andMilstein (1975) Nature 256:495-497, which discusses one method ofgenerating monoclonal antibodies. Summarized briefly, this methodinvolves injecting an animal with an immunogen to initiate a humoralimmune response. The animal is then sacrificed and cells taken from itsspleen, which are then fused with myeloma cells. The result is a hybridcell or "hybridoma" that is capable of reproducing in vitro. Thepopulation of hybridomas is then screened to isolate individual clones,each of which secretes a single antibody species to the immunogen. Inthis manner, the individual antibody species obtained are the productsof immortalized and cloned single B cells from the immune animalgenerated in response to a specific site recognized on the immunogenicsubstance.

Other suitable techniques involve selection of libraries of antibodiesin phage or similar vectors. See, Huse, et al. (1989) "Generation of aLarge Combinatorial Library of the Immunoglobulin Repertoire in PhageLambda," Science 246:1275-1281; and Ward, et al. (1989) Nature341:544-546. The polypeptides and antibodies of the present inventionmay be used with or without modification, including chimeric orhumanized antibodies. Frequently, the polypeptides and antibodies willbe labeled by joining, either covalently or non-covalently, a substancewhich provides for a detectable signal. A wide variety of labels andconjugation techniques are known and are reported extensively in boththe scientific and patent literature. Suitable labels includeradionuclides, enzymes, substrates, cofactors, inhibitors, fluorescentmoieties, chemiluminescent moieties, magnetic particles, and the like.Patents, teaching the use of such labels include U.S. Pat. Nos.3,817,837; 3,850,752; 3,939,350; 3,996,345; 4,277,437; 4,275,149; and4,366,241. Also, recombinant immunoglobulins may be produced. See,Cabilly, U.S. Pat. No. 4,816,567; and Queen, et al. (1989) Proc. Nat'lAcad, Sci. USA 86:10029-10033.

The antibodies of this invention can also be used for affinitychromatography in isolating each monocyte protein. Columns can beprepared where the antibodies are linked to a solid support, e.g.,particles, such as agarose, SEPHADEX, or the like, where a cell lysatemay be passed through the column, the column washed, followed byincreasing concentrations of a mild denaturant, whereby purifiedmonocyte protein will be released.

The antibodies may also be used to screen expression libraries forparticular expression products. Usually the antibodies used in such aprocedure will be labeled with a moiety allowing easy detection ofpresence of antigen by antibody binding.

Antibodies to monocyte proteins may be used for the analysis or, oridentification of specific cell population components which express therespective protein. By assaying the expression products of cellsexpressing monocyte proteins it is possible to diagnose disease, e.g.,immune-compromised conditions, monocyte depleted conditions, oroverproduction of monocyte.

Antibodies raised against each monocyte will also be useful to raiseanti-idiotypic antibodies. These will be useful in detecting ordiagnosing various immunological conditions related to expression of therespective antigens.

b. Immunoassays

A particular protein can be measured by a variety of immunoassaymethods. For a review of immunological and immunoassay procedures ingeneral, see Stites and Terr (eds.) 1991 Basic and Clinical Immunology(7th ed.). Moreover, the immunoassays of the present invention can beperformed in any of several configurations, which are reviewedextensively in Maggio (ed.) (1980) Enzyme Immunoassay CRC Press, BocaRaton, Fla.; Tijan (1985) "Practice and Theory of Enzyme Immunoassays,"Laboratory Techniques in Biochemistry and Molecular Biology, ElsevierScience Publishers B.V., Amsterdam; and Harlow and Lane Antibodies, ALaboratory Manual, supra, each of which is incorporated herein byreference. See also Chan (ed.) (1987) Immunoassay: A Practical GuideAcademic Press, Orlando, Fla.; Price and Newman (eds.) (1991) Principlesand Practice of Imunoassays Stockton Press, NY; and Ngo (ed.) (1988)Non-isotopic Immunoassays Plenum Press, NY.

Immunoassays for measurement of these monocyte proteins can be performedby a variety of methods known to those skilled in the art. In brief,immunoassays to measure the protein can be competitive or noncompetitivebinding assays. In competitive binding assays, the sample to be analyzedcompetes with a labeled analyte for specific binding sites on a captureagent bound to a solid surface. Preferably the capture agent is anantibody specifically reactive with the monocyte protein produced asdescribed above. The concentration of labeled analyte bound to thecapture agent is inversely proportional to the amount of free analytepresent in the sample.

In a competitive binding immunoassay, the monocyte protein present inthe sample competes with labeled protein for binding to a specificbinding agent, for example, an antibody specifically reactive with themonocyte protein. The binding agent may be bound to a solid surface toeffect separation of bound labeled protein from the unbound labeledprotein. Alternately, the competitive binding assay may be conducted inliquid phase and any of a variety of techniques known in the art may beused to separate the bound labeled protein from the unbound labeledprotein. Following separation, the amount of bound labeled protein isdetermined. The amount of protein present in the sample is inverselyproportional to the amount of labeled protein binding.

Alternatively, a homogeneous immunoassay may be performed in which aseparation step is not needed. In these immunoassays, the label on theprotein is altered by the binding of the protein to its specific bindingagent. This alteration in the labeled protein results in a decrease orincrease in the signal emitted by label, so that measurement of thelabel at the end of the immunoassay allows for detection or quantitationof the protein.

These monocyte proteins may also be quantitatively determined by avariety of noncompetitive immunoassay methods. For example, a two-site,solid phase sandwich immunoassay may be used. In this type of assay, abinding agent for the protein, for example an antibody, is attached to asolid support. A second protein binding agent, which may also be anantibody, and which binds the protein at a different site, is labeled.After binding at both sites on the protein has occurred, the unboundlabeled binding agent is removed and the amount of labeled binding agentbound to the solid phase is measured. The amount of labeled bindingagent bound is directly proportional to the amount of protein in thesample.

Western blot analysis can be used to determine the presence of monocyteproteins in a sample. Electrophoresis is carried out, e.g., on a tissuesample suspected of containing the protein. Following electrophoresis toseparate the proteins, and transfer of the proteins to a suitable solidsupport such as a nitrocellulose filter, the solid support is incubatedwith an antibody reactive with the denatured protein. This antibody maybe labeled, or alternatively may be it may be detected by subsequentincubation with a second labeled antibody that binds the primaryantibody.

The immunoassay formats described above employ labeled assay components.The label can be in a variety of forms. The label may be coupleddirectly or indirectly to the desired component of the assay accordingto methods well known in the art. A wide variety of labels may be used.The component may be labeled by any one of several methods.Traditionally a radioactive label incorporating ³ H, ¹²⁵ I, ³⁵ S, ¹⁴ C,or ³² P is used. Non-radioactive labels include ligands which bind tolabeled antibodies, fluorophores, chemiluminescent agents, enzymes, andantibodies which can serve as specific binding pair members for alabeled protein. The choice of label depends on sensitivity required,ease of conjugation with the compound, stability requirements, andavailable instrumentation. For a review of various labeling or signalproducing systems which may be used, see U.S. Pat. No. 4,391,904, whichis incorporated herein by reference.

Antibodies reactive with a particular protein can also be measured by avariety of immunoassay methods. For reviews of immunological andimmunoassay procedures applicable to the measurement of antibodies byimmunoassay techniques, see, e.g., Stites and Terr (eds.) Basic andClinical Immunology (7th ed.) supra; Maggio (ed.) Enzyme Immunoassay,supra; and Harlow and Lane Antibodies, A Laboratory Manual, supra.

A variety of different immunoassay formats, separation techniques, andlabels can be also be used similar to those described above for themeasurement of specific proteins. Moreover, many methods are known forevaluating selectivity of binding for specific protein or closelyrelated proteins.

VI. Purified monocyte proteins

The human monocyte FDF03 protein amino acid sequence is provided in SEQID NO: 2. Partial mouse sequence is provided in SEQ ID NO: 4. Human YE01amino acid and nucleotide sequences for the Ig-family member areprovided in SEQ ID NO: 5-10. The receptor family members, designatedKTE03, including the YYB01, YYB04, and KLM63, KLM66, and KLM67embodiments, are described in SEQ ID NO: 11-22.

The peptide sequences allow preparation of peptides to generateantibodies to recognize such segments, and allow preparation ofoligonucleotides which encode such sequences. Moreover, affinityreagents allow detection and purification of more protein, includingfull length or recombinant forms. And oligonucleotide sequences allowdetection of cDNAs encoding, or closely related to, these.

VII. Physical Variants

This invention also encompasses proteins or peptides having substantialamino acid sequence similarity with an amino acid sequence of SEQ ID NO:2 or 4; 6, 8, or 10; or 12, 14, 16, 18, 20, or 22, especially splicevariants. Variants exhibiting substitutions, e.g., 20 or fewer,preferably 10 or fewer, and more preferably 5 or fewer substitutions,are also enabled. Where the substitutions are conservativesubstitutions, the variants will share immunogenic or antigenicsimilarity or cross-reactivity with a corresponding natural sequenceprotein. Natural variants include individual, allelic, polymorphic,strain, or species variants.

Amino acid sequence similarity, or sequence identity, is determined byoptimizing residue matches, if necessary, by introducing gaps asrequired. This changes when considering conservative substitutions asmatches. Conservative substitutions typically include substitutionswithin the following groups: glycine, alanine; valine, isoleucine,leucine; aspartic acid, glutamic acid; asparagine, glutamine; serine,threonine; lysine, arginine; and phenylalanine, tyrosine. Homologousamino acid sequences include natural allelic and interspecies variationsin each respective protein sequence. Typical homologous proteins orpeptides will have from 50-100% similarity (if gaps can be introduced),to 75-100% similarity (if conservative substitutions are included) withthe amino acid sequence of the relevant monocyte protein. Identitymeasures will be at least about 50%, generally at least 60%, moregenerally at least 65%, usually at least 70%, more usually at least 75%,preferably at least 80%, and more preferably at least 80%, and inparticularly preferred embodiments, at least 85% or more. See alsoNeedleham, et al. (1970) J. Mol. Biol. 48:443-453; Sankoff, et al.(1983) Time Warps, String Edits, and Macromolecules: The Theory andPractice of Sequence Comparison Chapter One, Addison-Wesley, Reading,Mass.; and software packages from IntelliGenetics, Mountain View,Calif.; and the University of Wisconsin Genetics Computer Group (GCG),Madison, Wis.

Nucleic acids encoding the corresponding mammalian monocyte proteinswill typically hybridize, e.g., to SEQ ID NO 1 and/or 3; 5, 7, and/or 9;or 11, 13, 15, 17, 19, and/or 21 under stringent conditions. Forexample, nucleic acids encoding the respective monocyte proteins willtypically hybridize to the appropriate nucleic acid under stringenthybridization conditions, while providing few false positivehybridization signals. Generally, stringent conditions are selected tobe about 10° C. lower than the thermal melting point (Tm) for thesequence being hybridized to at a defined ionic strength and pH. The Tmis the temperature (under defined ionic strength and pH) at which 50% ofthe target sequence hybridizes to a perfectly matched probe. Typically,stringent conditions will be those in which the salt concentration inwash is about 0.02 molar at pH 7 and the temperature is at least about50° C. Other factors may significantly affect the stringency ofhybridization, including, among others, base composition and size of thecomplementary strands, the presence of organic solvents such asformamide, and the extent of base mismatching. A preferred embodimentwill include nucleic acids which will bind to disclosed sequences in 50%formamide and 20-50 mM NaCl at 42° C. In certain cases, the stringencymay be relaxed to detect other nucleic acids exhibiting less thancomplete sequence identity.

An isolated monocyte gene DNA can be readily modified by nucleotidesubstitutions, nucleotide deletions, nucleotide insertions, andinversions of nucleotide stretches. These modifications result in novelDNA sequences which encode these monocyte antigens, their derivatives,or proteins having highly similar physiological, immunogenic, orantigenic activity.

Modified sequences can be used to produce mutant antigens or to enhanceexpression. Enhanced expression may involve gene amplification,increased transcription, increased translation, and other mechanisms.Such mutant monocyte protein derivatives include predetermined orsite-specific mutations of the respective protein or its fragments."Mutant monocyte protein" encompasses a polypeptide otherwise fallingwithin the homology definition of the monocyte protein as set forthabove, but having an amino acid sequence which differs from that of themonocyte protein as found in nature, whether by way of deletion,substitution, or insertion. In particular, "site specific mutantmonocyte protein" generally includes proteins having significantsimilarity with a protein having a sequence of SEQ ID NO: 2 or 4; 6, 8,or 10; or 12, 14, 16, 18, 20, or 22. Generally, the variant will sharemany physicochemical and biological activities, e.g., antigenic orimmunogenic, with those sequences, and in preferred embodiments containmost or all of the disclosed sequence. Similar concepts apply to thesevarious monocyte proteins, particularly those found in various warmblooded animals, e.g., primates and mammals.

Although site specific mutation sites are predetermined, mutants neednot be site specific. Monocyte protein mutagenesis can be conducted bymaking amino acid insertions or deletions. Substitutions, deletions,insertions, or any combinations may be generated to arrive at a finalconstruct. Insertions include amino- or carboxyl-terminal fusions.Random mutagenesis can be conducted at a target codon and the expressedmutants can then be screened for the desired activity. Methods formaking substitution mutations at predetermined sites in DNA having aknown sequence are well known in the art, e.g., by M13 primermutagenesis or polymerase chain reaction (PCR) techniques. See also,Sambrook, et al. (1989) and Ausubel, et al. (1987 and Supplements). Themutations in the DNA normally should not place coding sequences out ofreading frames and preferably will not create complementary regions thatcould hybridize to produce secondary mRNA structure such as loops orhairpins.

The present invention also provides recombinant proteins, e.g.,heterologous fusion proteins using segments from these proteins. Aheterologous fusion protein is a fusion of proteins or segments whichare naturally not normally fused in the same manner. Thus, the fusionproduct of an immunoglobulin with a respective monocyte polypeptide is acontinuous protein molecule having sequences fused in a typical peptidelinkage, typically made as a single translation product and exhibitingproperties derived from each source peptide. A similar concept appliesto heterologous nucleic acid sequences.

In addition, new constructs may be made from combining similarfunctional domains from other proteins. For example, domains or othersegments may be "swapped" between different new fusion polypeptides orfragments, typically with related proteins, e.g., within the Ig familyor the Fc receptor family. Preferably, intact structural domains will beused, e.g., intact Ig portions. See, e.g., Cunningham, et al. (1989)Science 243:1330-1336; and O'Dowd, et al. (1988) J. Biol. Chem.263:15985-15992. Thus, new chimeric polypeptides exhibiting newcombinations of specificities will result from the functional linkage ofprotein-binding specificities and other functional domains. Also,alanine scanning mutagenesis may be applied, preferably to residueswhich structurally are exterior to the secondary structure, which willavoid most of the critical residues which generally disrupt tertiarystructure.

"Derivatives" of these monocyte antigens include amino acid sequencemutants, glycosylation variants, and covalent or aggregate conjugateswith other chemical moieties. Covalent derivatives can be prepared bylinkage of functionalities to groups which are found in these monocyteprotein amino acid side chains or at the N- or C-termini, by means whichare well known in the art. These derivatives can include, withoutlimitation, aliphatic esters or amides of the carboxyl terminus, or ofresidues containing carboxyl side chains, O-acyl derivatives of hydroxylgroup-containing residues, and N-acyl derivatives of the amino terminalamino acid or amino-group containing residues, e.g., lysine or arginine.Acyl groups are selected from the group of alkyl-moieties including C3to C18 normal alkyl, thereby forming alkanoyl aroyl species. Covalentattachment to carrier proteins may be important when immunogenicmoieties are haptens.

In particular, glycosylation alterations are included, e.g., made bymodifying the glycosylation patterns of a polypeptide during itssynthesis and processing, or in further processing steps. Particularlypreferred means for accomplishing this are by exposing the polypeptideto glycosylating enzymes derived from cells which normally provide suchprocessing, e.g., mammalian glycosylation enzymes. Deglycosylationenzymes are also contemplated. Also embraced are versions of the sameprimary amino acid sequence which have other minor modifications,including phosphorylated amino acid residues, e.g., phosphotyrosine,phosphoserine, or phosphothreonine, or other moieties, including ribosylgroups or cross-linking reagents. Also, proteins comprisingsubstitutions are encompassed, which should retain substantialimmunogenicity, to produce antibodies which recognize a protein of SEQID NO: 2 or 4; 6, 8, or 10; or 12, 14, 16, 18, 20, or 22. Alternatively,it may be desired to produce antibodies which recognize all or subsetsof SEQ ID NO: 2 and 4; 6, 8, and 10; or 12, 14, 16, 18, 20, and 22.Typically, these proteins will contain less than 20 residuesubstitutions from the disclosed sequence, more typically less than 10substitutions, preferably less than 5, and more preferably less than 3.Alternatively, proteins which begin and end at structural domains willusually retain antigenicity and cross immunogenicity.

A major group of derivatives are covalent conjugates of the monocyteproteins or fragments thereof with other proteins or polypeptides. Thesederivatives can be synthesized in recombinant culture such as N- orC-terminal fusions or by the use of agents known in the art for theirusefulness in cross-linking proteins through reactive side groups.Preferred protein derivatization sites with cross-linking agents are atfree amino groups, carbohydrate moieties, and cysteine residues.

Fusion polypeptides between these monocyte proteins and other homologousor heterologous proteins are also provided. Heterologous polypeptidesmay be fusions between different surface markers, resulting in, e.g., ahybrid protein. Likewise, heterologous fusions may be constructed whichwould exhibit a combination of properties or activities of thederivative proteins. Typical examples are fusions of a reporterpolypeptide, e.g., luciferase, with a segment or domain of a protein,e.g., a receptor-binding segment, so that the presence or location ofthe fused protein may be easily determined. See, e.g., Dull, et al.,U.S. Pat. No. 4,859,609. Other gene fusion partners include bacterialβ-galactosidase, trpE, Protein A, β-lactamase, alpha amylase, alcoholdehydrogenase, and yeast alpha mating factor. See, e.g., Godowski, etal. (1988) Science 241:812-816.

Such polypeptides may also have amino acid residues which have beenchemically modified by phosphorylation, sulfonation, biotinylation, orthe addition or removal of other moieties, particularly those which havemolecular shapes similar to phosphate groups. In some embodiments, themodifications will be useful labeling reagents, or serve as purificationtargets, e.g., affinity ligands.

This invention also contemplates the use of derivatives of thesemonocyte proteins other than variations in amino acid sequence orglycosylation. Such derivatives may involve covalent or aggregativeassociation with chemical moieties. These derivatives generally fallinto the three classes: (1) salts, (2) side chain and terminal residuecovalent modifications, and (3) adsorption complexes, for example withcell membranes. Such covalent or aggregative derivatives are useful asimmunogens, as reagents in immunoassays, or in purification methods suchas for affinity purification of ligands or other binding ligands. Forexample, a monocyte protein antigen can be immobilized by covalentbonding to a solid support such as cyanogen bromide-activated Sepharose,by methods which are well known in the art, or adsorbed onto polyolefinsurfaces, with or without glutaraldehyde cross-linking, for use in theassay or purification of anti-monocyte protein antibodies. The monocyteproteins can also be labeled with a detectable group, e.g.,radioiodinated by the chloramine T procedure, covalently bound to rareearth chelates, or conjugated to another fluorescent moiety for use indiagnostic assays. Purification of these monocyte proteins may beeffected by immobilized antibodies.

Isolated monocyte protein genes will allow transformation of cellslacking expression of a corresponding monocyte protein, e.g., eitherspecies types or cells which lack corresponding proteins and exhibitnegative background activity. Expression of transformed genes will allowisolation of antigenically pure cell lines, with defined or singlespecie variants. This approach will allow for more sensitive detectionand discrimination of the physiological effects of these monocyteproteins. Subcellular fragments, e.g., cytoplasts or membrane fragments,can be isolated and used.

VIII. Binding Agent:Monocyte Protein Complexes

A monocyte protein that specifically binds to or that is specificallyimmunoreactive with an antibody generated against a defined immunogen,such as an immunogen consisting of the amino acid sequence of SEQ ID NO:2 and/or 4; 6, 8, and/or 10; or 12, 14, 16, 18, 20, and/or 22, isdetermined in an immunoassay. The immunoassay uses a polyclonalantiserum which was raised to the protein of SEQ ID NO: 2, 4, 6, 8, 10,12, 14, 16, 18, 20, or 22, or appropriate combination. This antiserum isselected to have low crossreactivity against other members of therelated families, and any such crossreactivity is, or may be, removed byimmunoabsorption prior to use in the immunoassay.

In order to produce antisera for use in an immunoassay, the protein ofSEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, or 22 is isolated asdescribed herein. For example, recombinant protein may be produced in amammalian cell line. An inbred strain of mice such as Balb/c isimmunized with the appropriate protein using a standard adjuvant, suchas Freund's adjuvant, and a standard mouse immunization protocol (seeHarlow and Lane, supra). Alternatively, a synthetic peptide derived fromthe sequences disclosed herein and conjugated to a carrier protein canbe used an immunogen. Polyclonal sera are collected and titered againstthe immunogen protein in an immunoassay, e.g., a solid phase immunoassaywith the immunogen immobilized on a solid support. Polyclonal antiserawith a titer of 10⁴ or greater are selected and tested for their crossreactivity against other related proteins, using a competitive bindingimmunoassay such as the one described in Harlow and Lane, supra, atpages 570-573. See also Hertzenberg, et al. (eds. 1996) Weir's Handbookof Experimental Immunology vols. 1-4, Blackwell Science; and Coligan(1991) Current Protocols in Immunology Wiley/Greene, NY. Preferably twodifferent related proteins are used in this determination in conjunctionwith a given monocyte protein. For example, with the Ig family protein,at least two other family members are used to absorb out sharedepitopes. In conjunction with the Fc family member, two other members ofthe family are used. These other family members can be produced asrecombinant proteins and isolated using standard molecular biology andprotein chemistry techniques as described herein.

Immunoassays in the competitive binding format can be used for thecrossreactivity determinations. For example, the protein can beimmobilized to a solid support. Proteins added to the assay compete withthe binding of the antisera to the immobilized antigen. The ability ofthe above proteins to compete with the binding of the antisera to theimmobilized protein is compared to the protein of SEQ ID NO 2 and/or 4;6, 8, and/or 10; or 12, 14, 16, 18, 20, and/or 22. The percentcrossreactivity for the above proteins is calculated, using standardcalculations. Those antisera with less than 10% crossreactivity witheach of the proteins listed above are selected and pooled. Thecross-reacting antibodies are then removed from the pooled antisera byimmunoabsorption with the above-listed proteins.

The immunoabsorbed and pooled antisera are then used in a competitivebinding immunoassay as described above to compare a second protein tothe immunogen protein, e.g., the monocyte protein of SEQ ID NO: 2, 4, 6,8, 10, 12, 14, 16, 18, 20, or 22. In order to make this comparison, thetwo proteins are each assayed at a wide range of concentrations and theamount of each protein required to inhibit 50% of the binding of theantisera to the immobilized protein is determined. If the amount of thesecond protein required is less than twice the amount of the protein,e.g., of SEQ ID NO: 2, that is required, then the second protein is saidto specifically bind to an antibody generated to the immunogen.

It is understood that monocyte proteins are each a family of homologousproteins that comprise two or more genes. For a particular gene product,such as the human Ig family member protein, the invention encompassesnot only the amino acid sequences disclosed herein, but also to otherproteins that are allelic, polymorphic, non-allelic, or speciesvariants. It is also understood that the term "human monocyte protein"includes nonnatural mutations introduced by deliberate mutation usingconventional recombinant technology such as single site mutation, or byexcising short sections of DNA encoding these proteins or splicevariants from the gene, or by substituting or adding small numbers ofnew amino acids. Such minor alterations must substantially maintain theimmunoidentity of the original molecule and/or its biological activity.Thus, these alterations include proteins that are specificallyimmunoreactive with a designated naturally occurring respective monocyteprotein, for example, the human monocyte protein exhibiting SEQ ID NO:4. Particular protein modifications considered minor would includeconservative substitution of amino acids with similar chemicalproperties, as described above for each protein family as a whole. Byaligning a protein optimally with the protein of SEQ ID NO 2 and 4; 6,8, and 10; or 12, 14, 16, 18, 20, and 22, and by using the conventionalimmunoassays described herein to determine immunoidentity, one candetermine the protein compositions of the invention.

IX. Uses

The present invention provides reagents which will find use indiagnostic applications as described elsewhere herein, e.g., in thegeneral description for developmental abnormalities, or below in thedescription of kits for diagnosis.

Monocyte genes, e.g., DNA or RNA may be used as a component in aforensic assay. For instance, the nucleotide sequences provided may belabeled using, e.g., ³² P or biotin and used to probe standardrestriction fragment polymorphism blots, providing a measurablecharacter to aid in distinguishing between individuals. Such probes maybe used in well-known forensic techniques such as geneticfingerprinting. In addition, nucleotide probes made from monocytesequences may be used in in situ assays to detect chromosomalabnormalities.

Antibodies and other binding agents directed towards monocyte proteinsor nucleic acids may be used to purify the corresponding monocyteprotein molecule. As described in the Examples below, antibodypurification of monocyte proteins is both possible and practicable.Antibodies and other binding agents may also be used in a diagnosticfashion to determine whether monocyte components are present in a tissuesample or cell population using well-known techniques described herein.The ability to attach a binding agent to a monocyte protein provides ameans to diagnose disorders associated with expression misregulation.Antibodies and other monocyte protein binding agents may also be usefulas histological markers. As described in the examples below, theexpression of each of these proteins is limited to specific tissuetypes. By directing a probe, such as an antibody or nucleic acid to therespective monocyte protein, it is possible to use the probe todistinguish tissue and cell types in situ or in vitro.

This invention also provides reagents which may exhibit significanttherapeutic value. The monocyte proteins (naturally occurring orrecombinant), fragments thereof, and antibodies thereto, along withcompounds identified as having binding affinity to the monocyte protein,may be useful in the treatment of conditions associated with abnormalphysiology or development, including abnormal proliferation, e.g.,cancerous conditions, or degenerative conditions. Abnormalproliferation, regeneration, degeneration, and atrophy may be modulatedby appropriate therapeutic treatment using the compositions providedherein. For example, a disease or disorder associated with abnormalexpression or abnormal signaling by a monocyte, e.g., as an antigenpresenting cell, is a target for an agonist or antagonist of theprotein. The proteins likely play a role in regulation or development ofhematopoietic cells, e.g., lymphoid cells, which affect immunologicalresponses, e.g., antigen presentation and the resulting effectorfunctions.

For example, the DX26 antibody shows that inhibitory antibodies will beuseful in modulating NK or T cell functions, e.g., killing. Suchmodulation will typically be a 20% effect, either increasing ordecreasing, e.g., the killing effect, but in preferred embodiments willhave a 30%, 40%, 50%, or more. Because the distribution is also inmonocytes, the molecule will probably also affect the regulation ofmonocyte mediated or initiated effector functions of the immune system,e.g., autoimmune responses, transplantation rejection, graft vs. hostdisease, inflammatory conditions, etc. These molecules may also affectelimination of neoplastic conditions, e.g., tumor rejection.

Other abnormal developmental conditions are known in cell types shown topossess monocyte protein mRNA by northern blot analysis. See Berkow(ed.) The Merck Manual of Diagnosis and Therapy, Merck & Co., Rahway,N.J.; and Thorn, et al. Harrison's Principles of Internal Medicine,McGraw-Hill, NY. Developmental or functional abnormalities, e.g., of theimmune system, cause significant medical abnormalities and conditionswhich may be susceptible to prevention or treatment using compositionsprovided herein.

Recombinant monocyte proteins or antibodies might be purified and thenadministered to a patient. These reagents can be combined fortherapeutic use with additional active or inert ingredients, e.g., inconventional pharmaceutically acceptable carriers or diluents, e.g.,immunogenic adjuvants, along with physiologically innocuous stabilizersand excipients. In particular, these may be useful in a vaccine context,where the antigen is combined with one of these therapeutic versions ofagonists or antagonists. These combinations can be sterile filtered andplaced into dosage forms as by lyophilization in dosage vials or storagein stabilized aqueous preparations. This invention also contemplates useof antibodies or binding fragments thereof, including forms which arenot complement binding.

Drug screening using antibodies or receptor or fragments thereof canidentify compounds having binding affinity to these monocyte proteins,including isolation of associated components. Subsequent biologicalassays can then be utilized to determine if the compound has intrinsicstimulating activity and is therefore a blocker or antagonist in that itblocks the activity of the protein. Likewise, a compound havingintrinsic stimulating activity might activate the cell through theprotein and is thus an agonist in that it simulates the cell. Thisinvention further contemplates the therapeutic use of antibodies to theproteins as antagonists.

The quantities of reagents necessary for effective therapy will dependupon many different factors, including means of administration, targetsite, physiological state of the patient, and other medicantsadministered. Thus, treatment dosages should be titrated to optimizesafety and efficacy. Typically, dosages used in vitro may provide usefulguidance in the amounts useful for in situ administration of thesereagents. Animal testing of effective doses for treatment of particulardisorders will provide further predictive indication of human dosage.Various considerations are described, e.g., in Gilman, et al. (eds.)(1990) Goodman and Gilman's: The Pharmacological Bases of Therapeutics(8th ed.) Pergamon Press; and (1990) Remington's Pharmaceutical Sciences(17th ed.) Mack Publishing Co., Easton, Pa. Methods for administrationare discussed therein and below, e.g., for oral, intravenous,intraperitoneal, or intramuscular administration, transdermal diffusion,and others. Pharmaceutically acceptable carriers will include water,saline, buffers, and other compounds described, e.g., in the MerckIndex, Merck & Co., Rahway, N.J. Dosage ranges would ordinarily beexpected to be in amounts lower than 1 mM concentrations, typically lessthan about 10 μM concentrations, usually less than about 100 nM,preferably less than about 10 pM (picomolar), and most preferably lessthan about 1 fM (femtomolar), with an appropriate carrier. Slow releaseformulations, or a slow release apparatus will often be utilized forcontinuous administration.

The monocyte proteins, fragments thereof, and antibodies to it or itsfragments, antagonists, and agonists, could be administered directly tothe host to be treated or, depending on the size of the compounds, itmay be desirable to conjugate them to carrier proteins such as ovalbuminor serum albumin prior to their administration. Therapeutic formulationsmay be administered in many conventional dosage formulations. While itis possible for the active ingredient to be administered alone, it ispreferable to present it as a pharmaceutical formulation. Formulationstypically comprise at least one active ingredient, as defined above,together with one or more acceptable carriers thereof. Each carriershould be both pharmaceutically and physiologically acceptable in thesense of being compatible with the other ingredients and not injuriousto the patient. Formulations include those suitable for oral, rectal,nasal, or parenteral (including subcutaneous, intramuscular, intravenousand intradermal) administration. The formulations may conveniently bepresented in unit dosage form and may be prepared by any methods wellknown in the art of pharmacy. See, e.g., Gilman, et al. (eds.) (1990)Goodman and Gilman's: The Pharmacological Bases of Therapeutics (8thed.) Pergamon Press; and (1990) Remington's Pharmaceutical Sciences(17th ed.) Mack Publishing Co., Easton, Pa.; Avis, et al. (eds.) (1993)Pharmaceutical Dosage Forms: Parenteral Medications Dekker, N.Y.;Lieberman, et al. (eds.) (1990) Pharmaceutical Dosage Forms: TabletsDekker, N.Y.; and Lieberman, et al. (eds.) (1990) Pharmaceutical DosageForms: Disperse Systems Dekker, N.Y. The therapy of this invention maybe combined with or used in association with other chemotherapeutic orchemopreventive agents.

Both the naturally occurring and the recombinant form of the monocyteproteins of this invention are particularly useful in kits and assaymethods which are capable of screening compounds for binding activity tothe proteins. Several methods of automating assays have been developedin recent years so as to permit screening of tens of thousands ofcompounds in a short period. See, e.g., Fodor, et al. (1991) Science251:767-773, and other descriptions of chemical diversity libraries,which describe means for testing of binding affinity by a plurality ofcompounds. The development of suitable assays can be greatly facilitatedby the availability of large amounts of purified, e.g., soluble versionsof, monocyte protein as provided by this invention.

For example, antagonists can often be found once the protein has beenstructurally defined. Testing of potential protein analogs is nowpossible upon the development of highly automated assay methods using apurified surface protein. In particular, new agonists and antagonistswill be discovered by using screening techniques described herein. Ofparticular importance are compounds found to have a combined bindingaffinity for multiple related cell surface antigens, e.g., compoundswhich can serve as antagonists for species variants of a monocyteprotein.

This invention is particularly useful for screening compounds by usingrecombinant monocyte protein in a variety of drug screening techniques.The advantages of using a recombinant protein in screening for specificligands include: (a) improved renewable source of the protein from aspecific source; (b) potentially greater number of antigens per cellgiving better signal to noise ratio in assays; and (c) species variantspecificity (theoretically giving greater biological and diseasespecificity).

One method of drug screening utilizes eukaryotic or prokaryotic hostcells which are stably transformed with recombinant DNA moleculesexpressing a monocyte protein. Cells may be isolated which express thatprotein in isolation from any others. Such cells, either in viable orfixed form, can be used for standard surface protein binding assays. Seealso, Parce, et al. (1989) Science 246:243-247; and Owicki, et al.(1990) Proc. Nat'l Acad. Sci. USA 87:4007-4011, which describe sensitivemethods to detect cellular responses. Competitive assays areparticularly useful, where the cells (source of monocyte protein) arecontacted and incubated with an antibody having known binding affinityto the antigen, such as ¹²⁵ I-antibody, and a test sample whose bindingaffinity to the binding composition is being measured. The bound andfree labeled binding compositions are then separated to assess thedegree of protein binding. The amount of test compound bound isinversely proportional to the amount of labeled antibody binding to theknown source. Many techniques can be used to separate bound from freereagent to assess the degree of binding. This separation step couldtypically involve a procedure such as adhesion to filters followed bywashing, adhesion to plastic followed by washing, or centrifugation ofthe cell membranes. Viable cells could also be used to screen for theeffects of drugs on these monocyte protein mediated functions, e.g.,antigen presentation or helper function.

Another method utilizes membranes from transformed eukaryotic orprokaryotic host cells as the source of a monocyte protein. These cellsare stably transformed with DNA vectors directing the expression of theappropriate protein, e.g., an engineered membrane bound form.Essentially, the membranes would be prepared from the cells and used inbinding assays such as the competitive assay set forth above.

Still another approach is to use solubilized, unpurified or solubilized,purified monocyte protein from transformed eukaryotic or prokaryotichost cells. This allows for a "molecular" binding assay with theadvantages of increased specificity, the ability to automate, and highdrug test throughput.

Another technique for drug screening involves an approach which provideshigh throughput screening for compounds having suitable binding affinityto the respective monocyte protein and is described in detail in Geysen,European Patent Application 84/03564, published on Sep. 13, 1984. First,large numbers of different small peptide test compounds are synthesizedon a solid substrate, e.g., plastic pins or some other appropriatesurface, see Fodor, et al., supra. Then all the pins are reacted withsolubilized, unpurified or solubilized, purified monocyte protein, andwashed. The next step involves detecting bound reagent, e.g., antibody.

One means for determining which sites interact with specific otherproteins is a physical structure determination, e.g., x-raycrystallography or 2 dimensional NMR techniques. These will provideguidance as to which amino acid residues form molecular contact regions.For a detailed description of protein structural determination, see,e.g., Blundell and Johnson (1976) Protein Crystallography AcademicPress, NY.

X. Kits

This invention also contemplates use of these monocyte proteins,fragments thereof, peptides, and their fusion products in a variety ofdiagnostic kits and methods for detecting the presence of a monocyteprotein or message. Typically the kit will have a compartment containingeither a defined monocyte peptide or gene segment or a reagent whichrecognizes one or the other, e.g., antibodies.

A kit for determining the binding affinity of a test compound to therespective monocyte protein would typically comprise a test compound; alabeled compound, for example an antibody having known binding affinityfor the protein; a source of the monocyte protein (naturally occurringor recombinant); and a means for separating bound from free labeledcompound, such as a solid phase for immobilizing the monocyte protein.Once compounds are screened, those having suitable binding affinity tothe protein can be evaluated in suitable biological assays, as are wellknown in the art, to determine whether they act as agonists orantagonists to regulate monocyte function. The availability ofrecombinant monocyte polypeptides also provide well defined standardsfor calibrating such assays.

A preferred kit for determining the concentration of, for example, amonocyte protein in a sample would typically comprise a labeledcompound, e.g., antibody, having known binding affinity for the monocyteprotein, a source of monocyte protein (naturally occurring orrecombinant) and a means for separating the bound from free labeledcompound, for example, a solid phase for immobilizing the monocyteprotein. Compartments containing reagents, and instructions, willnormally be provided.

Antibodies, including antigen binding fragments, specific for therespective monocyte or its fragments are useful in diagnosticapplications to detect the presence of elevated levels of the proteinand/or its fragments. Such diagnostic assays can employ lysates, livecells, fixed cells, immunofluorescence, cell cultures, body fluids, andfurther can involve the detection of antigens in serum, or the like.Diagnostic assays may be homogeneous (without a separation step betweenfree reagent and antigen-monocyte protein complex) or heterogeneous(with a separation step). Various commercial assays exist, such asradioimmunoassay (RIA), enzyme-linked immunosorbent assay (ELISA),enzyme immunoassay (EIA), enzyme-multiplied immunoassay technique(EMIT), substrate-labeled fluorescent immunoassay (SLFIA), and the like.For example, unlabeled antibodies can be employed by using a secondantibody which is labeled and which recognizes the antibody to themonocyte protein or to a particular fragment thereof. Similar assayshave also been extensively discussed in the literature. See, e.g.,Harlow and Lane (1988) Antibodies: A Laboratory Manual, CSH Press, NY;Chan (ed.) (1987) Immunoassay: A Practical Guide Academic Press,Orlando, Fla.; Price and Newman (eds.) (1991) Principles and Practice ofImmunoassay Stockton Press, NY; and Ngo (ed.) (1988) NonisotopicImmunoassay Plenum Press, NY. In particular, the reagents may be usefulfor diagnosing monocyte populations in biological samples, either todetect an excess or deficiency of monocyte in a sample. The assay may bedirected to histological analysis of a biopsy, or evaluation of monocytenumbers in a blood or tissue sample.

Anti-idiotypic antibodies may have similar use to diagnose presence ofantibodies against a monocyte protein, as such may be diagnostic ofvarious abnormal states. For example, overproduction of the monocyteprotein may result in various immunological reactions which may bediagnostic of abnormal physiological states, particularly inproliferative cell conditions such as cancer or abnormaldifferentiation.

Frequently, the reagents for diagnostic assays are supplied in kits, soas to optimize the sensitivity of the assay. For the subject invention,depending upon the nature of the assay, the protocol, and the label,either labeled or unlabeled antibody or receptor, or labeled monocyteprotein is provided. This is usually in conjunction with otheradditives, such as buffers, stabilizers, materials necessary for signalproduction such as substrates for enzymes, and the like. Preferably, thekit will also contain instructions for proper use and disposal of thecontents after use. Typically the kit has compartments for each usefulreagent. Desirably, the reagents are provided as a dry lyophilizedpowder, where the reagents may be reconstituted in an aqueous mediumproviding appropriate concentrations of reagents for performing theassay.

Many of the aforementioned constituents of the drug screening and thediagnostic assays may be used without modification or may be modified ina variety of ways. For example, labeling may be achieved by covalentlyor non- covalently joining a moiety which directly or indirectlyprovides a detectable signal. In many of these assays, the protein, testcompound, monocyte protein, or antibodies thereto can be labeled eitherdirectly or indirectly. Possibilities for direct labeling include labelgroups: radiolabels such as ¹²⁵ I, enzymes (U.S. Pat. No. 3,645,090)such as peroxidase and alkaline phosphatase, and fluorescent labels(U.S. Pat. No. 3,940,475) capable of monitoring the change influorescence intensity, wavelength shift, or fluorescence polarization.Possibilities for indirect labeling include biotinylation of oneconstituent followed by binding to avidin coupled to one of the abovelabel groups.

There are also numerous methods of separating the bound from the freeprotein, or alternatively the bound from the free test compound. Themonocyte protein can be immobilized on various matrices followed bywashing. Suitable matrices include plastic such as an ELISA plate,filters, and beads. Methods of immobilizing the monocyte protein to amatrix include, without limitation, direct adhesion to plastic, use of acapture antibody, chemical coupling, and biotin-avidin. The last step inthis approach involves the precipitation of protein/antibody complex byone of several methods including those utilizing, e.g., an organicsolvent such as polyethylene glycol or a salt such as ammonium sulfate.Other suitable separation techniques include, without limitation, thefluorescein antibody magnetizable particle method described in Rattle,et al. (1984) Clin. Chem. 30:1457-1461, and the double antibody magneticparticle separation as described in U.S. Pat. No. 4,659,678.

Methods for linking proteins or their fragments to the various labelshave been extensively reported in the literature and do not requiredetailed discussion here. Many of the techniques involve the use ofactivated carboxyl groups either through the use of carbodiimide oractive esters to form peptide bonds, the formation of thioethers byreaction of a mercapto group with an activated halogen such aschloroacetyl, or an activated olefin such as maleimide, for linkage, orthe like. Fusion proteins will also find use in these applications.

Another diagnostic aspect of this invention involves use ofoligonucleotide or polynucleotide sequences taken from the sequence of arespective monocyte protein. These sequences can be used as probes fordetecting levels of the message in samples from patients suspected ofhaving an abnormal condition, e.g., cancer or immune problem. Thepreparation of both RNA and DNA nucleotide sequences, the labeling ofthe sequences, and the preferred size of the sequences has receivedample description and discussion in the literature. Normally anoligonucleotide probe should have at least about 14 nucleotides, usuallyat least about 18 nucleotides, and the polynucleotide probes may be upto several kilobases. Various labels may be employed, most commonlyradionuclides, particularly ³² P. However, other techniques may also beemployed, such as using biotin modified nucleotides for introductioninto a polynucleotide. The biotin then serves as the site for binding toavidin or antibodies, which may be labeled with a wide variety oflabels, such as radionuclides, fluorophores, enzymes, or the like.Alternatively, antibodies may be employed which can recognize specificduplexes, including DNA duplexes, RNA duplexes, DNA-RNA hybrid duplexes,or DNA-protein duplexes. The antibodies in turn may be labeled and theassay carried out where the duplex is bound to a surface, so that uponthe formation of duplex on the surface, the presence of antibody boundto the duplex can be detected. The use of probes to the novel anti-senseRNA may be carried out in any conventional techniques such as nucleicacid hybridization, plus and minus screening, recombinational probing,hybrid released translation (HRT), and hybrid arrested translation(HART). This also includes amplification techniques such as polymerasechain reaction (PCR).

Diagnostic kits which also test for the qualitative or quantitativepresence of other markers are also contemplated. Diagnosis or prognosismay depend on the combination of multiple indications used as markers.Thus, kits may test for combinations of markers. See, e.g., Viallet, etal. (1989) Progress in Growth Factor Res. 1:89-97.

XI. Binding Partner Isolation

Having isolated one member of a binding partner of a specificinteraction, methods exist for isolating the counter-partner. See,Gearing, et al. (1989) EMBO J. 8:3667-3676. For example, means to labela monocyte surface protein without interfering with the binding to itsreceptor can be determined. For example, an affinity label can be fusedto either the amino- or carboxyl-terminus of the ligand. An expressionlibrary can be screened for specific binding to the monocyte protein,e.g., by cell sorting, or other screening to detect subpopulations whichexpress such a binding component. See, e.g., Ho, et al. (1993) Proc.Nat'l Acad. Sci. USA 90:11267-11271. Alternatively, a panning method maybe used. See, e.g., Seed and Aruffo (1987) Proc. Nat'l Acad. Sci. USA84:3365-3369. A two-hybrid selection system may also be applied makingappropriate constructs with the available monocyte protein sequences.See, e.g., Fields and Song (1989) Nature 340:245-246.

Protein cross-linking techniques with label can be applied to isolatebinding partners of a monocyte protein. This would allow identificationof proteins which specifically interact with the appropriate monocyteprotein.

The broad scope of this invention is best understood with reference tothe following examples, which are not intended to limit the invention tospecific embodiments.

EXAMPLES

I. General Methods

Many of the standard methods below are described or referenced, e.g., inManiatis, et al. (1982) Molecular Cloning, A Laboratory Manual ColdSpring Harbor Laboratory, Cold Spring Harbor Press, NY; Sambrook, et al.(1989) Molecular Cloning: A Laboratory Manual (2d ed.) Vols. 1-3, CSHPress, NY; Ausubel, et al., Biology Greene Publishing Associates,Brooklyn, N.Y.; or Ausubel, et al. (1987 and Supplements) CurrentProtocols in Molecular Biology Wiley/Greene, NY; Innis, et al. (eds.)(1990) PCR Protocols; A Guide to Methods and Applications AcademicPress, NY.

Methods for protein purification include such methods as ammoniumsulfate precipitation, column chromatography, electrophoresis,centrifugation, crystallization, and others. See, e.g., Ausubel, et al.(1987 and periodic supplements); Deutscher (1990) "Guide to ProteinPurification," Methods in Enzymology vol. 182, and other volumes in thisseries; Coligan, et al. (1996 and periodic Supplements) CurrentProtocols in Protein Science Wiley/Greene, NY; and manufacturer'sliterature on use of protein purification products, e.g., Pharmacia,Piscataway, N.J., or Bio-Rad, Richmond, Calif. Combination withrecombinant techniques allow fusion to appropriate segments, e.g., to aFLAG sequence or an equivalent which can be fused via aprotease-removable sequence. See, e.g., Hochuli (1989) ChemischeIndustrie 12:69-70; Hochuli (1990) "Purification of Recombinant Proteinswith Metal Chelate Absorbent" in Setlow (ed.) Genetic Engineering,Principle and Methods 12:87-98, Plenum Press, NY; and Crowe, et al.(1992) OIAexpress: The High Level Expression & Protein PurificationSystem QUIAGEN, Inc., Chatsworth, Calif.

Standard immunological techniques are described, e.g., in Hertzenberg,et al. (eds. 1996) Weir's Handbook of Experimental Immunology vols. 1-4,Blackwell Science; Coligan (1991) Current Protocols in ImmunologyWiley/Greene, NY; and Methods in Enzymology volumes. 70, 73, 74, 84, 92,93, 108, 116, 121, 132, 150, 162, and 163. See also, e.g., Paul (ed.)(1993) Fundamental Immunology (3d ed.) Raven Press, N.Y.

FACS analyses are described in Melamed, et al. (1990) Flow Cytometry andSorting Wiley-Liss, Inc., New York, N.Y.; Shapiro (1988) Practical FlowCytometry Liss, New York, N.Y.; and Robinson, et al. (1993) Handbook ofFlow Cytometry Methods Wiley-Liss, New York, N.Y.

II. Isolation of Human Monocytes

Healthy donors were subjected to a leukophoresis. Percoll gradients wereused to isolate mononuclear cells which were then subject to centrifugalelutriation. See, Figdor, et al. (1982) Blood 60:46-53; and Plas, et al.(1988) Expt'l. Hematol. 16:355-359. This highly enriched monocytefraction was cultured for 5-7 days in the presence of GM-CSF (800 U/ml)and IL-4 (500 U/ml), as described in Romani, et al (1994) J. Exp. Med.180:83-93; and Sallusto, et al (1994) J. Exp. Med. 179:1109-1118.

For making dendritic cells, human CD34+ cells were obtained as follows.See, e.g., Caux, et al. (1995) pages 1-5 in Banchereau and SchmittDendritic Cells in Fundamental and Clinical Immunology Plenum Press, NY.Peripheral or cord blood cells, sometimes CD34+ selected, were culturedin the presence of Stem Cell Factor (SCF), GM-CSF, and TNF-α inendotoxin free RPMI 1640 medium (GIBCO, Grand Island, N.Y.) supplementedwith 10% (v/v) heat-inactivated fetal bovine serum (FBS; FlowLaboratories, Irvine, Calif.), 10 mM HEPES, 2 mM L-glutamine, 5×10⁻⁵ M2-mercaptoethanol, penicillin (100 μg/ml). This is referred to ascomplete medium.

CD34+ cells were seeded for expansion in 25 to 75 cm² flasks (Corning,N.Y.) at 2×10⁴ cells/ml. Optimal conditions were maintained by splittingthese cultures at day 5 and 10 with medium containing fresh GM-CSF andTNF-α (cell concentration: 1-3×10⁵ cells/ml). In certain cases, cellswere FACS sorted for CD1a expression at about day 6.

In certain situations, cells were routinely collected after 12 days ofculture, eventually adherent cells were recovered using a 5 mM EDTAsolution. In other situations, the CD1a+ cells were activated byresuspension in complete medium at 5×10⁶ cells/ml and activated for theappropriate time (e.g., 1 or 6 h) with 1 μg/ml phorbol 12-myristate13-acetate (PMA, Sigma) and 100 ng/ml ionomycin (Calbiochem, La Jolla,Calif.). These cells were expanded for another 6 days, and RNA isolatedfor cDNA library preparation.

III. RNA Isolation and Library Construction

Total RNA is isolated using, e.g., the guanidine thiocyanate/CsClgradient procedure as described by Chirgwin, et al. (1978) Biochem.18:5294-5299.

Alternatively, poly(A)+ RNA is isolated using the OLIGOTEX mRNAisolation kit (QIAGEN). Double stranded cDNA are generated using, e.g.,the SUPERSCRIPT plasmid system (Gibco BRL, Gaithersburg, Md.) for cDNAsynthesis and plasmid cloning. The resulting double stranded cDNA isunidirectionally cloned, e.g., into pSport1 and transfected byelectroporation into ELECTROMAX DH10BTM Cells (Gibco BRL, Gaithersburg,Md.).

IV. Sequencing

DNA isolated from randomly picked clones, or after subtractivehybridization using unactivated cells, were subjected to nucleotidesequence analysis using standard techniques. A Taq DiDeoxy Terminatorcycle sequencing kit (Applied Biosystems, Foster City, Calif.) can beused. The labeled DNA fragments are separated using a DNA sequencing gelof an appropriate automated sequencer. Alternatively, the isolated cloneis sequenced as described, e.g., in Maniatis, et al. (1982) MolecularCloning, A Laboratory Manual, Cold Spring Harbor Laboratory, Cold SpringHarbor Press; Sambrook, et al. (1989) Molecular Cloning: A LaboratoryManual, (2d ed.), vols. 1-3, CSH Press, NY; Ausubel, et al., Biology,Greene Publishing Associates, Brooklyn, N.Y.; or Ausubel, et al. (1987and Supplements) Current Protocols in Molecular Biology, Greene/Wiley,New York. Chemical sequencing methods are also available, e.g., usingMaxam and Gilbert sequencing techniques.

V. Isolation of Human Monocyte Protein Genes

The FDF03, the YE01, and KTE03 (YYB01 and YYB04) clones were sequenced,and analyzed for open reading frames. The clones were further analyzedto extend the nucleic acid sequence to a full, or nearly full, openreading frame.

mRNA is prepared from appropriate cell populations by the FastTrack kit(Invitrogen) from which cDNA is generated using, e.g., SuperscriptPlasmid System for cDNA synthesis from GIBCO-BRL (Gaithersburg, Md.)essentially as described by the manufacturer. Modification to theprocedure may include the substitution of other cloning adapters for theSal1 adapters provided with the kit. The resultant cDNA from these cellsis used to generate libraries, e.g., in the plasmid PcDNA II(Invitrogen). The cDNA is cloned into the polylinker and is used totransform an appropriate strain, e.g., DH10B, of E. coli. Plasmid isisolated and purified, e.g., with the Qiagen system (Chatsworth, Calif.)which is used to generate RNA probes from, e.g., the SP6 promoter.

RNA probes are labeled, e.g., using the Genius System(Boehringer-Mannheim) as described by the manufacturer. Filter lifts ofthe cDNA library can be pre-hybridized, e.g., at 42° C. for 3-6 hours inChurch's buffer (50% formamide, 6×SSPE, 50 mM NaHPO₄ pH 7.2, 7% SDS,0.1% N-Lauryl sarcosine, 2% Boehringer-Mannheim blocking reagent).Filters are probed, e.g., overnight in the same buffer containing theappropriate probes. The filters are washed, e.g., as described by theGenius System. The colonies that hybridize are selected.

The entire cDNA of human monocyte proteins are sequenced, e.g., by thedideoxynucleotide chain termination method with T7 polymerase (U.S.Biochemicals, Cleveland, Ohio) using double-stranded DNA as template.Data base searching and sequence analysis are performed usingIntelliGenetics programs (Mountain View, Calif.) to determine ifhomology exists between previously reported clones.

Table 1 discloses sequence encoding a human FDF03 gene and mousecounterpart sequence, and also shows alignment of available sequence.Likewise, Table 2 discloses three sequences encoding human YE01 geneproducts, including a splice variant and a transcript which encodes asoluble product. Table 3 provides sequences of embodiments of the KTE03gene products, and shows evidence of splice variants.

VI. Recombinant Monocyte Gene Constructs

Poly(A)⁺ RNA is isolated from appropriate cell populations, e.g., usingthe FastTrack mRNA kit (Invitrogen, San Diego, Calif.). Samples areelectrophoresed, e.g., in a 1% agarose gel containing formaldehyde andtransferred to a GeneScreen membrane (NEN Research Products, Boston,Mass.). Hybridization is performed, e.g., at 65° C. in 0.5 M NaHPO₄ pH7.2, 7% SDS, 1 mM EDTA, and 1% BSA (fraction V) with ³² P-dCTP labeledmonocyte gene cDNA at 10⁷ cpm/ml. After hybridization filters are washedthree times at 50° C. in 0.2×SSC, 0.1% SDS, and exposed to film for 24h.

The recombinant gene construct may be used to generate probe fordetecting the message. The insert may be excised and used in thedetection methods described above.

VII. Expression of Monocyte Gene Protein in E. coli.

PCR is used to make a construct comprising the open reading frame,preferably in operable association with proper promoter, selection, andregulatory sequences. The resulting expression plasmid is transformedinto an appropriate, e.g., the Topp5, E. coli strain (Stratagene, LaJolla, Calif.). Ampicillin resistant (50 μg/ml) transformants are grownin Luria Broth (Gibco) at 37° C. until the optical density at 550 nm is0.7. Recombinant protein is induced with 0.4 mMisopropyl-βD-thiogalactopyranoside (Sigma, St. Louis, Mo.) andincubation of the cells continued at 20° C. for a further 18 hours.Cells from a 1 liter culture are harvested by centrifugation andresuspended, e.g., in 200 ml of ice cold 30% sucrose, 50 mM Tris HCl pH8.0, 1 mM ethylenediamine-tetraacetic acid. After 10 min on ice, icecold water is added to a total volume of 2 liters. After 20 min on ice,cells are removed by centrifugation and the supernatant is clarified byfiltration via a 5 μM Millipak 60 (Millipore Corp., Bedford, Mass.).

The recombinant protein is purified via standard purification methods,e.g., various ion exchange chromatography methods. Immunoaffinitymethods using antibodies described below can also be used. Affinitymethods may be used where an epitope tag is engineered into anexpression construct.

VIII. Mapping of Human Monocyte Genes

DNA isolation, restriction enzyme digestion, agarose gelelectrophoresis, Southern blot transfer and hybridization are performedaccording to standard techniques. See Jenkins, et al. (1982) J. Virol.43:26-36. Blots may be prepared with Hybond-N nylon membrane (Amersham).The probe is labeled with ³² P-dCTP; washing is done to a finalstringency, e.g., of 0.1×SSC, 0.1% SDS, 65° C.

Alternatively, a BIOS Laboratories (New Haven, Conn.) mouse somatic cellhybrid panel may be combined with PCR methods.

IX. Analysis of Individual Variation

From the distribution data, an abundant easily accessible cell type isselected for sampling from individuals. Using PCR techniques, a largepopulation of individuals are analyzed for this gene. cDNA or other PCRmethods are used to sequence the corresponding gene in the differentindividuals, and their sequences are compared. This indicates both theextent of divergence among racial or other populations, as well asdetermining which residues are likely to be modifiable without dramaticeffects on function.

X. Preparation of Antibodies

Recombinant monocyte proteins are generated by expression in E. coli asshown above, and tested for biological activity. Active or denaturedproteins may be used for immunization of appropriate mammals for eitherpolyclonal serum production, or for monoclonal antibody production.Antibodies are selected for use in Western blots, against native ordenatured antigen, and for those which modulate a biological activity.

Antibodies prepared against the FDF03 have confirmed specific binding ondendritic cells.

XI. Isolation of Counterpart Primate Monocyte Genes

Human cDNA clones encoding these genes are used as probes, or to designPCR primers to find counterparts in various primate species, e.g.,chimpanzees.

XII. Use of Reagents to Analyze Cell Populations

Detection of the level of monocyte cells present in a sample isimportant for diagnosis of certain aberrant disease conditions. Forexample, an increase in the number of monocytes in a tissue or the lymphsystem can be indicative of the presence of a monocyte hyperplasia,tissue or graft rejection, or inflammation. A low monocyte populationcan indicate an abnormal reaction to, e.g., a bacterial or viralinfection, which may require the appropriate treat to normalize themonocyte response.

FACS analysis using a labeled binding agent specific for a cell surfacemonocyte protein, see, e.g., Melamed, et al. (1990) Flow Cytometry andSorting Wiley-Liss, Inc., New York, N.Y.; Shapiro (1988) Practical FlowCytometry Liss, New York, N.Y.; and Robinson, et al. (1993) Handbook ofFlow Cytometry Methods Wiley-Liss, New York, N.Y., is used indetermining the number of monocytes present in a cell mixture, e.g.,PBMCs, adherent cells, etc. The binding agent is also used forhistological analysis of tissue samples, either fresh or fixed, toanalyze infiltration of monocyte. Diverse cell populations may also beevaluated, either in a cell destructive assay, or in certain assayswhere cells retain viability.

Analysis of the presence of soluble intracellular molecules isperformed, e.g., with a fluorescent binding agent specific for amonocyte as described in Openshaw, et al. (1995) J. Exp, Med.182:1357-1367. alternatively, tissue or cell fixation methods may beused.

Levels of monocyte transcripts are quantitated, e.g., usingsemiquantitative PCR as described in Murphy, et al. (1993) J. Immunol.Methods 162:211-223. Primers are designed such that genomic DNA is notdetected.

Distribution of the FDF03 embodiment has been studied usinghybridization and PCR analysis. Northern blot analysis locatedtranscripts in dendritic cells and the JY cell line. There appear to betwo transcripts of about 700 bp and 1300 bp, which may be differentiallyregulated, and an estimated frequency of about 1 in 4000 in restingmonocytes or LPS and IFNγ activated monocytes. the shorter message doesnot appear to encode a soluble version of the protein, e.g., lacking theTM and intracellular segments. Southern blot analysis has detectedtranscripts in monocytes, dendritic cells, PBMC, B cells, and splenic Bcells. The message appears to be down-regulated upon monocyteactivation.

Distribution of the YE01 embodiment has also been evaluated. The messageappears to be monocyte specific, and is a low abundance message. It isdetectable in cDNA Southern blots in resting monocytes, and in activatedmonocytes. Its highest expression was found in 6 hour LPS stimulatedmonocytes. It is also detectable in anti-CD3 and PMA activated PBMC. Itmay be faintly detectable in dendritic cells, but this may be due tocontamination of the dendritic cell population with residual monocytes.At that level of sensitivity, it is undetectable in NK cells, B or Tcells, or any fetal cells examined. However, the YE01 gene product isspecifically recognized by a monoclonal antibody DX26. This antibody,when cross-linked, can inhibit NK cell mediated killing of certaintargets. The antibody recognizes protein expressed in T cells, B cells,NK cells, and monocytes. The gene encoding the antigen recognized byDX26, which is apparently a polymorphic variant of the YE01 isolate, hasbeen cloned and has essentially the sequence:

The KTE03 expression levels were also investigated. The message appearedto be up-regulated upon IL-10 exposure when the monocytes were activatedby LPS and IFNγ.

XIII. Isolation of a Binding Counterpart

A monocyte protein can be used as a specific binding reagent, by takingadvantage of its specificity of binding, much like an antibody would beused. A binding reagent is either labeled as described above, e.g.,fluorescence or otherwise, or immobilized to a substrate for panningmethods.

The monocyte protein is used to screen for a cell line which exhibitsbinding. Standard staining techniques are used to detect or sortintracellular or surface expressed ligand, or surface expressingtransformed cells are screened by panning. Screening of intracellularexpression is performed by various staining or immunofluorescenceprocedures. See also McMahan, et al. (1991) EMBO J. 10:2821-2832.

For example, on day 0, precoat 2-chamber permanox slides with 1 ml perchamber of fibronectin, 10 ng/ml in PBS, for 30 min at room temperature.Rinse once with PBS. Then plate COS cells at 2-3×10⁵ cells per chamberin 1.5 ml of growth media. Incubate overnight at 37° C.

On day 1 for each sample, prepare 0.5 ml of a solution of 66 mg/mlDEAE-dextran, 66 mM chloroquine, and 4 mg DNA in serum free DME. Foreach set, a positive control is prepared, e.g., of human receptor-FLAGcDNA at 1 and 1/200 dilution, and a negative mock. Rinse cells withserum free DME. Add the DNA solution and incubate 5 hr at 37° C. Removethe medium and add 0.5 ml 10% DMSO in DYE for 2.5 min. Remove and washonce with DME. Add 1.5 ml growth medium and incubate overnight.

On day 2, change the medium. On days 3 or 4, the cells are fixed andstained. Rinse the cells twice with Hank's Buffered Saline Solution(HBSS) and fix in 4% paraformaldehyde (PFA)/glucose for 5 min. Wash 3×with HBSS. The slides may be stored at -80° C. after all liquid isremoved. For each chamber, 0.5 ml incubations are performed as follows.Add HBSS/saponin(0.1%) with 32 ml/ml of 1M NaN₃ for 20 min. Cells arethen washed with HBSS/saponin 1×. Add protein or protein/antibodycomplex to cells and incubate for 30 min. Wash cells twice withHBSS/saponin. If appropriate, add first antibody for 30 min. Add secondantibody, e.g., Vector anti-mouse antibody, at 1/200 dilution, andincubate for 30 min. Prepare ELISA solution, e.g., Vector Elite ABChorseradish peroxidase solution, and preincubate for 30 min. Use, e.g.,1 drop of solution A (avidin) and 1 drop solution B (biotin) per 2.5 mlHBSS/saponin. Wash cells twice with HBSS/saponin. Add ABC HRP solutionand incubate for 30 min. Wash cells twice with HBSS, second wash for 2min, which closes cells. Then add Vector diaminobenzoic acid (DAB) for 5to 10 min. Use 2 drops of buffer plus 4 drops DAB plus 2 drops of H₂ O₂per 5 ml of glass distilled water. Carefully remove chamber and rinseslide in water. Air dry for a few minutes, then add 1 drop of CrystalMount and a cover slip. Bake for 5 min at 85-90° C.

Alternatively, other monocyte protein specific binding reagents are usedto affinity purify or sort out cells expressing a receptor. See, e.g.,Sambrook, et al. or Ausubel, et al.

Another strategy is to screen for a membrane bound receptor by panning.The receptor cDNA is constructed as described above. The ligand can beimmobilized and used to immobilize expressing cells. Immobilization maybe achieved by use of appropriate antibodies which recognize, e.g., aFLAG sequence of a monocyte protein fusion construct, or by use ofantibodies raised against the first antibodies. Recursive cycles ofselection and amplification lead to enrichment of appropriate clones andeventual isolation of ligand expressing clones.

Phage expression libraries can be screened by monocyte protein.Appropriate label techniques, e.g., anti-FLAG antibodies, will allowspecific labeling of appropriate clones.

XIV. Isolation of a Soluble YE01

An additional family member of the previously described YE01, alsodesignated DNAX Leukocyte Associated Immunoglobulin-like Receptor(DLAIR; and now designated DLAIR-1) was cloned by screening a human Tcell tumor line cDNA library (TcT). Bacterial colony lift membranes werehybridized with a DLAIR-1 probe comprising a BglII-SphI digestionfragment, spanning the Ig loop in the extracellular domain. Twopositives were isolated and sequenced. Sequence analysis revealed thatboth clones contained identical open reading frames of 414 base pairs,encoding a 135 amino acid protein with a predicted 21 amino acid leadersequence and a predicted molecular weight of 14.7 kDa. This molecule,now referred to as DLAIR-2, contains one Ig loop. See Table 2. The Igloop has 84% homology with DLAIR-1, indicating that it belongs to thesame family, but is encoded by a separate gene. DLAIR-2 lacks atransmembrane region which suggests that it is a secreted protein.

DLAIR-2, as a soluble molecule with similarity to DLAIR-1, may be usedas an antagonist to this inhibitory receptor.

XV. Preparation of DX26 Monoclonal Antibody

Mice were immunized with a human NK cell clone and antibodies werescreened for their capacity to inhibit NK cell-mediated lysis of FcRbearing targets. Alternatively, antibodies will be raised to purifiedprotein.

XVI. Cross-linking DLAIR-1 With mAb Inhibits NK Cell-mediated Killing

DX26 mAb did not inhibit NK clone killing of the HLA-negativeEBV-transformed B cell line 721.221. However, when 721.221 wastransfected with the human FcγR-II (CD32) and used as a target, NKcell-mediated cytolysis was inhibited by DX26 mAb. This indicates thatsignaling through the molecule recognized by DX26 mkb (designated DNAXLeukocyte Associated Immunoglobulin-like Receptor (DLAIR)), delivers anegative signal to NK cell clones that prevents their killing specifictarget cells. In agreement with this, NK cell-mediated cytotoxicityagainst Colo-205, PA-1, or FO-1, each an FcR-negative human cell line,was not inhibited by the addition of DX26 mAb. Moreover, lysis of P815cells, an FcR-expressing mouse mastocytoma cell line, which is killed invitro by human NK cell clones upon simultaneous cross linking of CD2,CD16, CD69, or DNAM-1 antigen, was also inhibited by DX26 mAb. Theseresults lead to a conclusion that DLAIR delivers a strong inhibitorysignal to NK cells, since the positive signal given by potent inducersof NK cell cytotoxicity was overruled by DX26 mAb.

XVII. DLAIR-1 is an Inhibitory Receptor On Resting NSK cells

NK cell clones consist of clonally derived populations of activated NKcells. These cells are potently inhibited by DLAIR signaling. We set outto study whether DLAIR is also functioning as an inhibitory receptor onNK cells that had not been previously activated. Resting NK cells,prepared from peripheral blood by negative depletion using magneticbeads, were able to lyse P815 target cells when simultaneously activatedthrough CD16. This MS cell mediated cytotoxicity was inhibited by theaddition of DX26 mAb. Thus, DLAIR is functional as an inhibitoryreceptor on both activated and resting NK cells.

XVIII. DLAIR is a Widely Expressed Antigen

Phenotypic analysis of human peripheral blood lymphocytes demonstratedthat DLAIR is a widely distributed molecule. In healthy donor PBMC, CD3⁺CD4⁺ T cells (70-80%), CD3⁺ CD8⁺ T cells (80-90%), CD3⁻ CD56⁺ NK cells(95-100%), CD3⁻ CD19⁺ B cells (80-90%), and CD3⁻ CD14+ monocytes(99-100%) all expressed the DLAIR molecule. Human fetal thymocytes, boththe immature CD4⁺ CD8⁺ cells and mature CD4⁺ CD8⁻ or CD4⁻ CD8⁺ singlepositive cells also expressed DLAIR. Peripheral blood granulocytes,platelets and erythrocytes did not express DLAIR.

Human NK cell clones and T cell clones all expressed DLAIR, with theexception of the long-term cultured NK clones NKL and NK92 (see Table4). EBV-transformed B cell lines, the B cell tumor Daudi, and the NKtumor cell line YT and several non-hematopoietic cell lines did notexpress DLAIR, whereas human T cell lines did show DLAIR expression.

                       TABLE 4                                                

    ______________________________________                                        Expression of DLAIR on human tumor cell lines.sup.1                                                        control IgGl                                                                  DX26 mAb                                         cell line                                                                              type         (mean fluorescence intensity)                           ______________________________________                                         HUT78   T cell tumor <5         25.8                                           Peer           T cell tumor              <5            29.1                   Molt4          T cell tumor               <5            30.7                  CEM            T cell tumor              <5            92.7                   Jurkat         T cell tumor              <5            47.1                   HL60           promyeloid tumor         <5            46.9                    U937           myeloid tumor           <5            49.5                     721.221        EBV-B cell            <5     <5                                JY             EBV-B cell            <5     <5                                Daudi          B cell tumor          <5     <5                                YT             NK cell tumor         <5     <5                                NKL            NK cell clone         <5      <5                               NK92           NK cell clone         <5      <5                               Colo205        colon carcinorna      <5      <5                               293T           embryonic kidney      <5     <5                                PA-1           teratocarcinoma       <5     <5                                FO-1           melanoma              <5    <5                               ______________________________________                                         .sup.1 cells were stained with control IgGl or DX26 mAb and PEconjugated      goatanti-mouse-IgG as a second step.     Cells   were analyzed on a           FACScan.                                                                 

XIX. Expression Cloning of the DX26 Antigen

The DX26 antibody was used to expression clone the antigen the antibodyrecognizes. The expression cloning was performed using standard methods.See, e.g., Sambrook, et al. or Coligan, et al.

DX26 antigen is expression cloned, e.g., from a polyclonal humanactivated NK cell cDNA library in the pJFE14 expression vector. COS7cells are transfected with the library and antigen positive cells wereselected using phycoerythrin labeled anti-DX26 mAb. The cDNA sequencewas determined and found to match much of the YE01 sequence. The DX26antibody specifically binds to the product of the YE01 gene product.

In another method, oligonucleotides are used to screen a library. Incombination with polymerase chain reaction (PCR) techniques, syntheticoligonucleotides in appropriate orientations are used as primers toselect correct clones from a library.

Moreover, the YE01 gene product is specifically recognized by amonoclonal antibody DX26. This antibody, when crosslinked, can inhibitNK cell mediated killing of certain targets. The antibody recognizesprotein expressed in T cells, B cells, NK cells, and monocytes. The geneencoding the antigen recognized by DX26, which is apparently apolymorphic variant of the YE01 isolate, has been cloned and hasessentially the sequence (see SEQ ID NO: 7). This isolate has adifferent 3' untranslated sequence from the original YE01 transcript,apparently due to use of an alternative polyadenylation site. A solubleform of DLAIR has also been detected (see SEQ ID NO: 9).

Distribution analysis of the DX26 isolate has determined, Northern blotanalysis, the distribution as follows. Probing of mRNA of human NK cellclones with DLAIR cDNA, PBMC, the human T cell line Jurkat, and thehuman myeloid cell line Jurkat results in two bands of approximately1800 bp and 3000-4000 bp. This indicates that besides the cloned cDNA,another transcript with sequence similarity to DLAIR exists in thesecell lines. Whether this contains the same open reading frame is atpresent unknown, but will be determined upon cloning and sequenceanalysis of that transcript. The EBV-transformed human B cell line JYdid not show transcripts that probed with DLAIR cDNA.

XX. DLAIR-1 Binds SHP-1 and SHP-2

The existence of two consensus sequences for ITIMs within thecytoplasmic domain of DLAIR-1, suggested that the generation of aninhibitory signal in NK cells was manifested by the recruitment of SHP-1and/or SHP-2. To determine if DLAIR-1 was capable of binding proteintyrosine phosphatases, a NK cell clone was stimulated with pervanadate(an inhibitor of protein tyrosine phosphatases that induces tyrosinephosphorylation (O'Shea, et al. (1992) Proc. Natl. Acad. Sci. USA89:10306-10310), lysed, and immunoprecipitated with DX26 MAb.Immunoprecipitates were then analyzed by Western blot using antibodiesspecific for SHP-1 and SHP-2. Both SHP-1 and SHP-2 associated withtyrosine phosphorylated DLAIR-1. These results suggest that recruitmentof SHP-1 and SHP-2 may be involved in mediating the negative signaltransduced via engagement of the DLAIR-1 molecule.

All references cited herein are incorporated herein by reference to thesame extent as if each individual publication or patent application wasspecifically and individually indicated to be incorporated by referencein its entirety for all purposes.

Many modifications and variations of this invention can be made withoutdeparting from its spirit and scope, as will be apparent to thoseskilled in the art. The specific embodiments described herein areoffered by way of example only, and the invention is to be limited onlyby the terms of the appended claims, along with the full scope ofequivalents to which such claims are entitled.

    __________________________________________________________________________    #             SEQUENCE LISTING                                                   - -  - - (1) GENERAL INFORMATION:                                             - -    (iii) NUMBER OF SEQUENCES: 22                                          - -  - - (2) INFORMATION FOR SEQ ID NO:1:                                     - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 1249 base - #pairs                                                (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 154..1062                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: mat.sub.-- - #peptide                                           (B) LOCATION: 211..1062                                              - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:1:                               - - GTTTGGGGAA GGCTCCTGGC CCCCACAGCC CTCTTCGGAG CCTGAGCCCG GC -            #TCTCCTCA     60                                                                 - - CTCACCTCAA CCCCCAGGCG GCCCCTCCAC AGGGCCCCTC TCCTGCCTGG AC -            #GGCTCTGC    120                                                                 - - TGGTCTCCCC GTCCCCTGGA GAAGAACAAG GCC ATG GGT CGG CC - #C CTG CTG       CTG     174                                                                                       - #                  - # Met Gly Arg Pro Leu Leu Leu                         - #                  - # -19             -15                 - - CCC CTA CTG CCC CTG CTG CTG CCG CCA GCA TT - #T CTG CAG CCT AGT GGC          222                                                                       Pro Leu Leu Pro Leu Leu Leu Pro Pro Ala Ph - #e Leu Gln Pro Ser Gly                   -10          - #        -5          - #         1                      - - TCC ACA GGA TCT GGT CCA AGC TAC CTT TAT GG - #G GTC ACT CAA CCA AAA          270                                                                       Ser Thr Gly Ser Gly Pro Ser Tyr Leu Tyr Gl - #y Val Thr Gln Pro Lys             5                - #  10                - #  15                - #  20       - - CAC CTC TCA GCC TCC ATG GGT GGC TCT GTG GA - #A ATC CCC TTC TCC TTC          318                                                                       His Leu Ser Ala Ser Met Gly Gly Ser Val Gl - #u Ile Pro Phe Ser Phe                            25 - #                 30 - #                 35              - - TAT TAC CCC TGG GAG TTA GCC ACA GCT CCC GA - #C GTG AGA ATA TCC TGG          366                                                                       Tyr Tyr Pro Trp Glu Leu Ala Thr Ala Pro As - #p Val Arg Ile Ser Trp                        40     - #             45     - #             50                  - - AGA CGG GGC CAC TTC CAC GGG CAG TCC TTC TA - #C AGC ACA AGG CCG CCT          414                                                                       Arg Arg Gly His Phe His Gly Gln Ser Phe Ty - #r Ser Thr Arg Pro Pro                    55         - #         60         - #         65                      - - TCC ATT CAC AAG GAT TAT GTG AAC CGG CTC TT - #T CTG AAC TGG ACA GAG          462                                                                       Ser Ile His Lys Asp Tyr Val Asn Arg Leu Ph - #e Leu Asn Trp Thr Glu                70             - #     75             - #     80                          - - GGT CAG AAG AGC GGC TTC CTC AGG ATC TCC AA - #C CTG CAG AAG CAG GAC          510                                                                       Gly Gln Lys Ser Gly Phe Leu Arg Ile Ser As - #n Leu Gln Lys Gln Asp            85                 - # 90                 - # 95                 - #100       - - CAG TCT GTG TAT TTC TGC CGA GTT GAG CTG GA - #C ACA CGG AGC TCA GGG          558                                                                       Gln Ser Val Tyr Phe Cys Arg Val Glu Leu As - #p Thr Arg Ser Ser Gly                           105  - #               110  - #               115              - - AGG CAG CAG TGG CAG TCC ATC GAG GGG ACC AA - #A CTC TCC ATC ACC CAG          606                                                                       Arg Gln Gln Trp Gln Ser Ile Glu Gly Thr Ly - #s Leu Ser Ile Thr Gln                       120      - #           125      - #           130                  - - GCT GTC ACG ACC ACC ACC CAG AGG CCC AGC AG - #C ATG ACT ACC ACC TGG          654                                                                       Ala Val Thr Thr Thr Thr Gln Arg Pro Ser Se - #r Met Thr Thr Thr Trp                   135          - #       140          - #       145                      - - AGG CTC AGT AGC ACA ACC ACC ACA ACC GGC CT - #C AGG GTC ACA CAG GGC          702                                                                       Arg Leu Ser Ser Thr Thr Thr Thr Thr Gly Le - #u Arg Val Thr Gln Gly               150              - #   155              - #   160                          - - AAA CGA CGC TCA GAC TCT TGG CAC ATA AGT CT - #G GAG ACT GCT GTG GGG          750                                                                       Lys Arg Arg Ser Asp Ser Trp His Ile Ser Le - #u Glu Thr Ala Val Gly           165                 1 - #70                 1 - #75                 1 -      #80                                                                              - - GTG GCA GTG GCT GTC ACT GTG CTC GGA ATC AT - #G ATT TTG GGA CTG        ATC      798                                                                    Val Ala Val Ala Val Thr Val Leu Gly Ile Me - #t Ile Leu Gly Leu Ile                          185  - #               190  - #               195              - - TGC CTC CTC AGG TGG AGG AGA AGG AAA GGT CA - #G CAG CGG ACT AAA GCC          846                                                                       Cys Leu Leu Arg Trp Arg Arg Arg Lys Gly Gl - #n Gln Arg Thr Lys Ala                       200      - #           205      - #           210                  - - ACA ACC CCA GCC AGG GAA CCC TTC CAA AAC AC - #A GAG GAG CCA TAT GAG          894                                                                       Thr Thr Pro Ala Arg Glu Pro Phe Gln Asn Th - #r Glu Glu Pro Tyr Glu                   215          - #       220          - #       225                      - - AAT ATC AGG AAT GAA GGA CAA AAT ACA GAT CC - #C AAG CTA AAT CCC AAG          942                                                                       Asn Ile Arg Asn Glu Gly Gln Asn Thr Asp Pr - #o Lys Leu Asn Pro Lys               230              - #   235              - #   240                          - - GAT GAC GGC ATC GTA TAT GCT TCC CTT GCC CT - #C TCC AGC TCC ACC TCA          990                                                                       Asp Asp Gly Ile Val Tyr Ala Ser Leu Ala Le - #u Ser Ser Ser Thr Ser           245                 2 - #50                 2 - #55                 2 -      #60                                                                              - - CCC AGA GCA CCT CCC AGC CAC CGT CCC CTC AA - #G AGC CCC CAG AAC        GAG     1038                                                                    Pro Arg Ala Pro Pro Ser His Arg Pro Leu Ly - #s Ser Pro Gln Asn Glu                          265  - #               270  - #               275              - - ACC CTG TAC TCT GTC TTA AAG GCC TAACCAATGG AC - #AGCCCTCT CAAGACTGAA        1092                                                                       Thr Leu Tyr Ser Val Leu Lys Ala                                                           280                                                                - - TGGTGAGGCC AGGTACAGTG GCGCACACCT GTAATCCCAG CTACTCTGAA GC -             #CTGAGGCA   1152                                                                 - - GAATCAAGTG AGCCCAGGAG TTCAGGGCCA GCTTTGATAA TGGAGCGAGA TG -            #CCATCTCT   1212                                                                 - - AGTTAAAAAT ATATATTAAC AATAAAGTAA CAAATTT      - #                      - #    1249                                                                     - -  - - (2) INFORMATION FOR SEQ ID NO:2:                                     - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 303 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:2:                               - - Met Gly Arg Pro Leu Leu Leu Pro Leu Leu Pr - #o Leu Leu Leu Pro Pro     19             -15    - #             -10    - #              -5                - - Ala Phe Leu Gln Pro Ser Gly Ser Thr Gly Se - #r Gly Pro Ser Tyr Leu                    1    - #           5       - #           10                      - - Tyr Gly Val Thr Gln Pro Lys His Leu Ser Al - #a Ser Met Gly Gly Ser           15             - #     20             - #     25                          - - Val Glu Ile Pro Phe Ser Phe Tyr Tyr Pro Tr - #p Glu Leu Ala Thr Ala       30                 - # 35                 - # 40                 - # 45       - - Pro Asp Val Arg Ile Ser Trp Arg Arg Gly Hi - #s Phe His Gly Gln Ser                       50 - #                 55 - #                 60              - - Phe Tyr Ser Thr Arg Pro Pro Ser Ile His Ly - #s Asp Tyr Val Asn Arg                   65     - #             70     - #             75                  - - Leu Phe Leu Asn Trp Thr Glu Gly Gln Lys Se - #r Gly Phe Leu Arg Ile               80         - #         85         - #         90                      - - Ser Asn Leu Gln Lys Gln Asp Gln Ser Val Ty - #r Phe Cys Arg Val Glu           95             - #    100             - #    105                          - - Leu Asp Thr Arg Ser Ser Gly Arg Gln Gln Tr - #p Gln Ser Ile Glu Gly      110                 1 - #15                 1 - #20                 1 -      #25                                                                              - - Thr Lys Leu Ser Ile Thr Gln Ala Val Thr Th - #r Thr Thr Gln Arg        Pro                                                                                             130  - #               135  - #               140             - - Ser Ser Met Thr Thr Thr Trp Arg Leu Ser Se - #r Thr Thr Thr Thr Thr                  145      - #           150      - #           155                  - - Gly Leu Arg Val Thr Gln Gly Lys Arg Arg Se - #r Asp Ser Trp His Ile              160          - #       165          - #       170                      - - Ser Leu Glu Thr Ala Val Gly Val Ala Val Al - #a Val Thr Val Leu Gly          175              - #   180              - #   185                          - - Ile Met Ile Leu Gly Leu Ile Cys Leu Leu Ar - #g Trp Arg Arg Arg Lys      190                 1 - #95                 2 - #00                 2 -      #05                                                                              - - Gly Gln Gln Arg Thr Lys Ala Thr Thr Pro Al - #a Arg Glu Pro Phe        Gln                                                                                             210  - #               215  - #               220             - - Asn Thr Glu Glu Pro Tyr Glu Asn Ile Arg As - #n Glu Gly Gln Asn Thr                  225      - #           230      - #           235                  - - Asp Pro Lys Leu Asn Pro Lys Asp Asp Gly Il - #e Val Tyr Ala Ser Leu              240          - #       245          - #       250                      - - Ala Leu Ser Ser Ser Thr Ser Pro Arg Ala Pr - #o Pro Ser His Arg Pro          255              - #   260              - #   265                          - - Leu Lys Ser Pro Gln Asn Glu Thr Leu Tyr Se - #r Val Leu Lys Ala          270                 2 - #75                 2 - #80                            - -  - - (2) INFORMATION FOR SEQ ID NO:3:                                     - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 376 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 78..374                                                - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:3:                               - - CCCCAGTGTC CCTAGACAGA GCATCCTTGC CTTCCTGATG GCTTTGCTGA TC -             #TCGCTTCC     60                                                                 - - CTGGAGGGAC TCCAGCC ATG GCT CAG GTC CTG CTT CTG - #CTC TCA TCA GGC           110                                                                                         - # Met Ala Gln Val Leu Leu Leu Leu Ser Ser - #Gly                            - #   1               - #5                  - #10            - - TGT CTG CAT GCT GGA AAT TCA GAA AGA TAC AA - #C AGA AAA AAT GGC TTT          158                                                                       Cys Leu His Ala Gly Asn Ser Glu Arg Tyr As - #n Arg Lys Asn Gly Phe                        15     - #             20     - #             25                  - - GGG GTC AAC CAA CCT GAA CGC TGC TCT GGA GT - #C CAG GGT GGC TCC ATC          206                                                                       Gly Val Asn Gln Pro Glu Arg Cys Ser Gly Va - #l Gln Gly Gly Ser Ile                    30         - #         35         - #         40                      - - GAC ATC CCC TTC TCC TTC TAT TTC CCC TGG AA - #G TTG GCC AAG GAT CCA          254                                                                       Asp Ile Pro Phe Ser Phe Tyr Phe Pro Trp Ly - #s Leu Ala Lys Asp Pro                45             - #     50             - #     55                          - - CAG ATG AGC ATA GCC TGG AAA TGG AAG GAT TT - #C CAT GGG GAA GTC ATC          302                                                                       Gln Met Ser Ile Ala Trp Lys Trp Lys Asp Ph - #e His Gly Glu Val Ile            60                 - # 65                 - # 70                 - # 75       - - TAC AAC TCC TCC CTG CCT TTC ATA CAT GAG CA - #C TTC AAG GGC CGG CTC          350                                                                       Tyr Asn Ser Ser Leu Pro Phe Ile His Glu Hi - #s Phe Lys Gly Arg Leu                            80 - #                 85 - #                 90              - - ATC CTG AAC TGG ACA CAG GGT CAG AC    - #                  - #                 376                                                                     Ile Leu Asn Trp Thr Gln Gly Gln                                                            95                                                                - -  - - (2) INFORMATION FOR SEQ ID NO:4:                                     - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 99 amino - #acids                                                 (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:4:                               - - Met Ala Gln Val Leu Leu Leu Leu Ser Ser Gl - #y Cys Leu His Ala Gly        1               5 - #                 10 - #                 15              - - Asn Ser Glu Arg Tyr Asn Arg Lys Asn Gly Ph - #e Gly Val Asn Gln Pro                   20     - #             25     - #             30                  - - Glu Arg Cys Ser Gly Val Gln Gly Gly Ser Il - #e Asp Ile Pro Phe Ser               35         - #         40         - #         45                      - - Phe Tyr Phe Pro Trp Lys Leu Ala Lys Asp Pr - #o Gln Met Ser Ile Ala           50             - #     55             - #     60                          - - Trp Lys Trp Lys Asp Phe His Gly Glu Val Il - #e Tyr Asn Ser Ser Leu       65                 - # 70                 - # 75                 - # 80       - - Pro Phe Ile His Glu His Phe Lys Gly Arg Le - #u Ile Leu Asn Trp Thr                       85 - #                 90 - #                 95              - - Gln Gly Gln                                                               - -  - - (2) INFORMATION FOR SEQ ID NO:5:                                     - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 1279 base - #pairs                                                (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 155..1015                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: misc.sub.-- - #feature                                          (B) LOCATION: 1247                                                            (D) OTHER INFORMATION: - #/note= "nucleotide 1247 designated                       C, but - #may be C or T"                                        - -     (ix) FEATURE:                                                                  (A) NAME/KEY: mat.sub.-- - #peptide                                           (B) LOCATION: 218..1015                                              - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:5:                               - - ACCGGTCCGG AATTCCCGGG TCGACCCACG CGTCCGGGAA GCCCCATAGG CA -             #GGAGGCCC     60                                                                 - - CCGGGCAGCA CATCCTGTCT GCTTGTGTCT GCTGCAGAGT TCTGTCCTTG CA -            #TTGGTGCG    120                                                                 - - CCTCAGGCCA GGCTGCACTG CTGGGACCTG GGCC ATG TCT CCC C - #AC CCC ACC            172                                                                                        - #                  - #  Met Ser Pro His Pro Thr                             - #                  - #  -21 -20                            - - GCC CTC CTG GGC CTA GTG CTC TGC CTG GCC CA - #G ACC ATC CAC ACG CAG          220                                                                       Ala Leu Leu Gly Leu Val Leu Cys Leu Ala Gl - #n Thr Ile His Thr Gln          15                 - - #10                  - #-5                  - # 1        - - GAG GAA GAT CTG CCC AGA CCC TCC ATC TCG GC - #T GAG CCA GGC ACC GTG          268                                                                       Glu Glu Asp Leu Pro Arg Pro Ser Ile Ser Al - #a Glu Pro Gly Thr Val                         5    - #              10    - #              15                  - - ATC CCC CTG GGG AGC CAT GTG ACT TTC GTG TG - #C CGG GGC CCG GTT GGG          316                                                                       Ile Pro Leu Gly Ser His Val Thr Phe Val Cy - #s Arg Gly Pro Val Gly                    20         - #         25         - #         30                      - - GTT CAA ACA TTC CGC CTG GAG AGG GAG AGT AG - #A TCC ACA TAC AAT GAT          364                                                                       Val Gln Thr Phe Arg Leu Glu Arg Glu Ser Ar - #g Ser Thr Tyr Asn Asp                35             - #     40             - #     45                          - - ACT GAA GAT GTG TCT CAA GCT AGT CCA TCT GA - #G TCA GAG GCC AGA TTC          412                                                                       Thr Glu Asp Val Ser Gln Ala Ser Pro Ser Gl - #u Ser Glu Ala Arg Phe            50                 - # 55                 - # 60                 - # 65       - - CGC ATT GAC TCA GTA AGT GAA GGA AAT GCC GG - #G CCT TAT CGC TGC ATC          460                                                                       Arg Ile Asp Ser Val Ser Glu Gly Asn Ala Gl - #y Pro Tyr Arg Cys Ile                            70 - #                 75 - #                 80              - - TAT TAT AAG CCC CCT AAA TGG TCT GAG CAG AG - #T GAC TAC CTG GAG CTG          508                                                                       Tyr Tyr Lys Pro Pro Lys Trp Ser Glu Gln Se - #r Asp Tyr Leu Glu Leu                        85     - #             90     - #             95                  - - CTG GTG AAA GAA ACC TCT GGA GGC CCG GAC TC - #C CCG GAC ACA GAG CCC          556                                                                       Leu Val Lys Glu Thr Ser Gly Gly Pro Asp Se - #r Pro Asp Thr Glu Pro                   100          - #       105          - #       110                      - - GGC TCC TCA GCT GGA CCC ACG CAG AGG CCG TC - #G GAC AAC AGT CAC AAT          604                                                                       Gly Ser Ser Ala Gly Pro Thr Gln Arg Pro Se - #r Asp Asn Ser His Asn               115              - #   120              - #   125                          - - GAG CAT GCA CCT GCT TCC CAA GGC CTG AAA GC - #T GAG CAT CTG TAT ATT          652                                                                       Glu His Ala Pro Ala Ser Gln Gly Leu Lys Al - #a Glu His Leu Tyr Ile           130                 1 - #35                 1 - #40                 1 -      #45                                                                              - - CTC ATC GGG GTC TCA GTG GTC TTC CTC TTC TG - #T CTC CTC CTC CTG        GTC      700                                                                    Leu Ile Gly Val Ser Val Val Phe Leu Phe Cy - #s Leu Leu Leu Leu Val                          150  - #               155  - #               160              - - CTC TTC TGC CTC CAT CGC CAG AAT CAG ATA AA - #G CAG GGG CCC CCC AGA          748                                                                       Leu Phe Cys Leu His Arg Gln Asn Gln Ile Ly - #s Gln Gly Pro Pro Arg                       165      - #           170      - #           175                  - - AGC AAG GAC GAG GAG CAG AAG CCA CAG CAG AG - #G CCT GAC CTG GCT GTT          796                                                                       Ser Lys Asp Glu Glu Gln Lys Pro Gln Gln Ar - #g Pro Asp Leu Ala Val                   180          - #       185          - #       190                      - - GAT GTT CTA GAG AGG ACA GCA GAC AAG GCC AC - #A GTC AAT GGA CTT CCT          844                                                                       Asp Val Leu Glu Arg Thr Ala Asp Lys Ala Th - #r Val Asn Gly Leu Pro               195              - #   200              - #   205                          - - GAG AAG GAC AGA GAG ACG GAC ACC TCG GCC CT - #G GCT GCA GGG AGT TCC          892                                                                       Glu Lys Asp Arg Glu Thr Asp Thr Ser Ala Le - #u Ala Ala Gly Ser Ser           210                 2 - #15                 2 - #20                 2 -      #25                                                                              - - CAG GAG GTG ACG TAT GCT CAG CTG GAC CAC TG - #G GCC CTC ACA CAG        AGG      940                                                                    Gln Glu Val Thr Tyr Ala Gln Leu Asp His Tr - #p Ala Leu Thr Gln Arg                          230  - #               235  - #               240              - - ACA GCC CGG GCT GTG TCC CCA CAG TCC ACA AA - #G CCC ATG GCC GAG TCC          988                                                                       Thr Ala Arg Ala Val Ser Pro Gln Ser Thr Ly - #s Pro Met Ala Glu Ser                       245      - #           250      - #           255                  - - ATC ACG TAT GCA GCC GTT GCC AGA CAC TGACCCCAT - #A CCCACCTGGC               1035                                                                       Ile Thr Tyr Ala Ala Val Ala Arg His                                                   260          - #       265                                             - - CTCTGCACCT GAGGGTAGAA AGTCACTCTA GGAAAAGCCT GAAGCAGCCA TT -             #TGGAAGGC   1095                                                                 - - TTCCTGTTGG ATTCCTCTTC ATCTAGAAAG CCAGCCAGGC AGCTGTCCTG GA -            #GACAAGAG   1155                                                                 - - CTGGAGACTG GAGGTTTCTA ACCAGCATCC AGAAGGTTCG TTAGCCAGGT GG -            #TCCCTTCT   1215                                                                 - - ACAATCGGAC AGCTCCTTGG ACAGACTGTT TCTCAGTTAT TTCCAAAAAC CC -            #AGCTACAG   1275                                                                 - - TTCC                 - #                  - #                  - #               1279                                                                  - -  - - (2) INFORMATION FOR SEQ ID NO:6:                                     - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 287 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:6:                               - - Met Ser Pro His Pro Thr Ala Leu Leu Gly Le - #u Val Leu Cys Leu Ala     21 -20                - # -15                - # -10                            - - Gln Thr Ile His Thr Gln Glu Glu Asp Leu Pr - #o Arg Pro Ser Ile Ser       -5                 - #  1               5 - #                 10              - - Ala Glu Pro Gly Thr Val Ile Pro Leu Gly Se - #r His Val Thr Phe Val                   15     - #             20     - #             25                  - - Cys Arg Gly Pro Val Gly Val Gln Thr Phe Ar - #g Leu Glu Arg Glu Ser               30         - #         35         - #         40                      - - Arg Ser Thr Tyr Asn Asp Thr Glu Asp Val Se - #r Gln Ala Ser Pro Ser           45             - #     50             - #     55                          - - Glu Ser Glu Ala Arg Phe Arg Ile Asp Ser Va - #l Ser Glu Gly Asn Ala       60                 - # 65                 - # 70                 - # 75       - - Gly Pro Tyr Arg Cys Ile Tyr Tyr Lys Pro Pr - #o Lys Trp Ser Glu Gln                       80 - #                 85 - #                 90              - - Ser Asp Tyr Leu Glu Leu Leu Val Lys Glu Th - #r Ser Gly Gly Pro Asp                   95     - #            100     - #            105                  - - Ser Pro Asp Thr Glu Pro Gly Ser Ser Ala Gl - #y Pro Thr Gln Arg Pro              110          - #       115          - #       120                      - - Ser Asp Asn Ser His Asn Glu His Ala Pro Al - #a Ser Gln Gly Leu Lys          125              - #   130              - #   135                          - - Ala Glu His Leu Tyr Ile Leu Ile Gly Val Se - #r Val Val Phe Leu Phe      140                 1 - #45                 1 - #50                 1 -      #55                                                                              - - Cys Leu Leu Leu Leu Val Leu Phe Cys Leu Hi - #s Arg Gln Asn Gln        Ile                                                                                             160  - #               165  - #               170             - - Lys Gln Gly Pro Pro Arg Ser Lys Asp Glu Gl - #u Gln Lys Pro Gln Gln                  175      - #           180      - #           185                  - - Arg Pro Asp Leu Ala Val Asp Val Leu Glu Ar - #g Thr Ala Asp Lys Ala              190          - #       195          - #       200                      - - Thr Val Asn Gly Leu Pro Glu Lys Asp Arg Gl - #u Thr Asp Thr Ser Ala          205              - #   210              - #   215                          - - Leu Ala Ala Gly Ser Ser Gln Glu Val Thr Ty - #r Ala Gln Leu Asp His      220                 2 - #25                 2 - #30                 2 -      #35                                                                              - - Trp Ala Leu Thr Gln Arg Thr Ala Arg Ala Va - #l Ser Pro Gln Ser        Thr                                                                                             240  - #               245  - #               250             - - Lys Pro Met Ala Glu Ser Ile Thr Tyr Ala Al - #a Val Ala Arg His                      255      - #           260      - #           265                  - -  - - (2) INFORMATION FOR SEQ ID NO:7:                                     - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 1728 base - #pairs                                                (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 69..929                                                - -     (ix) FEATURE:                                                                  (A) NAME/KEY: mat.sub.-- - #peptide                                           (B) LOCATION: 132..929                                               - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:7:                               - - AAAGGCTGCA GAGTTCTGTC CTTGCATTGG TGCGCCTCAG GCCAGGCTGC AC -             #TGCTGGGA     60                                                                 - - CCTGGGCC ATG TCT CCC CAC CCC ACC GCC CTC CTG - #GGC CTA GTG CTC        TGC     110                                                                              Met Ser Pro His Pro T - #hr Ala Leu Leu Gly Leu Val Leu Cys                  -21 -20       - #          -15       - #          -10                 - - CTG GCC CAG ACC ATC CAC ACG CAG GAG GAA GA - #T CTG CCC AGA CCC TCC          158                                                                       Leu Ala Gln Thr Ile His Thr Gln Glu Glu As - #p Leu Pro Arg Pro Ser                    -5         - #          1        - #       5                          - - ATC TCG GCT GAG CCA GGC ACC GTG ATC CCC CT - #G GGG AGC CAT GTG ACT          206                                                                       Ile Ser Ala Glu Pro Gly Thr Val Ile Pro Le - #u Gly Ser His Val Thr            10                 - # 15                 - # 20                 - # 25       - - TTC GTG TGC CGG GGC CCG GTT GGG GTT CAA AC - #A TTC CGC CTG GAG AGG          254                                                                       Phe Val Cys Arg Gly Pro Val Gly Val Gln Th - #r Phe Arg Leu Glu Arg                            30 - #                 35 - #                 40              - - GAG AGT AGA TCC ACA TAC AAT GAT ACT GAA GA - #T GTG TCT CAA GCT AGT          302                                                                       Glu Ser Arg Ser Thr Tyr Asn Asp Thr Glu As - #p Val Ser Gln Ala Ser                        45     - #             50     - #             55                  - - CCA TCT GAG TCA GAG GCC AGA TTC CGC ATT GA - #C TCA GTA AGT GAA GGA          350                                                                       Pro Ser Glu Ser Glu Ala Arg Phe Arg Ile As - #p Ser Val Ser Glu Gly                    60         - #         65         - #         70                      - - AAT GCC GGG CCT TAT CGC TGC ATC TAT TAT AA - #G CCC CCT AAA TGG TCT          398                                                                       Asn Ala Gly Pro Tyr Arg Cys Ile Tyr Tyr Ly - #s Pro Pro Lys Trp Ser                75             - #     80             - #     85                          - - GAG CAG AGT GAC TAC CTG GAG CTG CTG GTG AA - #A GAA ACC TCT GGA GGC          446                                                                       Glu Gln Ser Asp Tyr Leu Glu Leu Leu Val Ly - #s Glu Thr Ser Gly Gly            90                 - # 95                 - #100                 - #105       - - CCG GAC TCC CCG GAC ACA GAG CCC GGC TCC TC - #A GCT GGA CCC ACG CAG          494                                                                       Pro Asp Ser Pro Asp Thr Glu Pro Gly Ser Se - #r Ala Gly Pro Thr Gln                           110  - #               115  - #               120              - - AGG CCG TCG GAC AAC AGT CAC AAT GAG CAT GC - #A CCT GCT TCC CAA GGC          542                                                                       Arg Pro Ser Asp Asn Ser His Asn Glu His Al - #a Pro Ala Ser Gln Gly                       125      - #           130      - #           135                  - - CTG AAA GCT GAG CAT CTG TAT ATT CTC ATC GG - #G GTC TCA GTG GTC TTC          590                                                                       Leu Lys Ala Glu His Leu Tyr Ile Leu Ile Gl - #y Val Ser Val Val Phe                   140          - #       145          - #       150                      - - CTC TTC TGT CTC CTC CTC CTG GTC CTC TTC TG - #C CTC CAT CGC CAG AAT          638                                                                       Leu Phe Cys Leu Leu Leu Leu Val Leu Phe Cy - #s Leu His Arg Gln Asn               155              - #   160              - #   165                          - - CAG ATA AAG CAG GGG CCC CCC AGA AGC AAG GA - #C GAG GAG CAG AAG CCA          686                                                                       Gln Ile Lys Gln Gly Pro Pro Arg Ser Lys As - #p Glu Glu Gln Lys Pro           170                 1 - #75                 1 - #80                 1 -      #85                                                                              - - CAG CAG AGG CCT GAC CTG GCT GTT GAT GTT CT - #A GAG AGG ACA GCA        GAC      734                                                                    Gln Gln Arg Pro Asp Leu Ala Val Asp Val Le - #u Glu Arg Thr Ala Asp                          190  - #               195  - #               200              - - AAG GCC ACA GTC AAT GGA CTT CCT GAG AAG GA - #C AGA GAG ACG GAC ACC          782                                                                       Lys Ala Thr Val Asn Gly Leu Pro Glu Lys As - #p Arg Glu Thr Asp Thr                       205      - #           210      - #           215                  - - TCG GCC CTG GCT GCA GGG AGT TCC CAG GAG GT - #G ACG TAT GCT CAG CTG          830                                                                       Ser Ala Leu Ala Ala Gly Ser Ser Gln Glu Va - #l Thr Tyr Ala Gln Leu                   220          - #       225          - #       230                      - - GAC CAC TGG GCC CTC ACA CAG AGG ACA GCC CG - #G GCT GTG TCC CCA CAG          878                                                                       Asp His Trp Ala Leu Thr Gln Arg Thr Ala Ar - #g Ala Val Ser Pro Gln               235              - #   240              - #   245                          - - TCC ACA AAG CCC ATG GCC GAG TCC ATC ACG TA - #T GCA GCC GTT GCC AGA          926                                                                       Ser Thr Lys Pro Met Ala Glu Ser Ile Thr Ty - #r Ala Ala Val Ala Arg           250                 2 - #55                 2 - #60                 2 -      #65                                                                              - - CAC TGACCCCATA CCCACCTGGC CTCTGCACCT GAGGGTAGAA AGTCACTCT - #A               979                                                                      His                                                                            - - GGAAAAGCCT GAAGCAGCCA TTTGGAAGGC TTCCTGTTGG ATTCCTCTTC AT -             #CTAGAAAG   1039                                                                 - - CCAGCCAGGC AGCTGTCCTG GAGACAAGAG CTGGAGACTG GAGGTTTCTA AC -            #CAGCATCC   1099                                                                 - - AGAAGGTTCG TTAGCCAGGT GGTCCCTTCT ACAATCGAGC AGCTCCTTGG AC -            #AGACTGTT   1159                                                                 - - TCTCAGTTAT TTCCAGAGAC CCAGCTACAG TTCCCTGGCT GTTTCTAGAG AC -            #CCAGCTTT   1219                                                                 - - ATTCACCTGA CTGTTTCCAG AGACCCAGCT AAAGTCACCT GCCTGTTCTA AA -            #GGCCCAGC   1279                                                                 - - TACAGCCAAT CAGCCGATTT CCTGAGCAGT GATGCCACCT CCAAGCTTGT CC -            #TAGGTGTC   1339                                                                 - - TGCTGTGAAC CTCCAGTGAC CCCAGAGACT TTGCTGTAAT TATCTGCCCT GC -            #TGACCCTA   1399                                                                 - - AAGACCTTCC TAGAAGTCAA GAGCTAGCCT TGAGACTGTG CTATACACAC AC -            #AGCTGAGA   1459                                                                 - - GCCAAGCCCA GTTCTCTGGG TTGTGCTTTA CTCCACGCAT CAATAAATAA TT -            #TTGAAGGC   1519                                                                 - - CTCACATCTG GCAGCCCCAG GCCTGGTCCT GGGTGCATAG GTCTCTCGGA CC -            #CACTCTCT   1579                                                                 - - GCCTTCACAG TTGTTCAAAG CTGAGTGAGG GAAACAGGAC TTACGAAAAC GT -            #GTCAGCGT   1639                                                                 - - TTTCTTTTTA AAATTTAATT GATCAGGATT GTACGTAAAA AAAAAAAAAA AA -            #AAAAAAAA   1699                                                                 - - AAAAAAAAAA AAAAAAAAAA AAAAAAAGG         - #                  - #              1728                                                                     - -  - - (2) INFORMATION FOR SEQ ID NO:8:                                     - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 287 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:8:                               - - Met Ser Pro His Pro Thr Ala Leu Leu Gly Le - #u Val Leu Cys Leu Ala     21 -20                - # -15                - # -10                            - - Gln Thr Ile His Thr Gln Glu Glu Asp Leu Pr - #o Arg Pro Ser Ile Ser       -5                 - #  1               5 - #                 10              - - Ala Glu Pro Gly Thr Val Ile Pro Leu Gly Se - #r His Val Thr Phe Val                   15     - #             20     - #             25                  - - Cys Arg Gly Pro Val Gly Val Gln Thr Phe Ar - #g Leu Glu Arg Glu Ser               30         - #         35         - #         40                      - - Arg Ser Thr Tyr Asn Asp Thr Glu Asp Val Se - #r Gln Ala Ser Pro Ser           45             - #     50             - #     55                          - - Glu Ser Glu Ala Arg Phe Arg Ile Asp Ser Va - #l Ser Glu Gly Asn Ala       60                 - # 65                 - # 70                 - # 75       - - Gly Pro Tyr Arg Cys Ile Tyr Tyr Lys Pro Pr - #o Lys Trp Ser Glu Gln                       80 - #                 85 - #                 90              - - Ser Asp Tyr Leu Glu Leu Leu Val Lys Glu Th - #r Ser Gly Gly Pro Asp                   95     - #            100     - #            105                  - - Ser Pro Asp Thr Glu Pro Gly Ser Ser Ala Gl - #y Pro Thr Gln Arg Pro              110          - #       115          - #       120                      - - Ser Asp Asn Ser His Asn Glu His Ala Pro Al - #a Ser Gln Gly Leu Lys          125              - #   130              - #   135                          - - Ala Glu His Leu Tyr Ile Leu Ile Gly Val Se - #r Val Val Phe Leu Phe      140                 1 - #45                 1 - #50                 1 -      #55                                                                              - - Cys Leu Leu Leu Leu Val Leu Phe Cys Leu Hi - #s Arg Gln Asn Gln        Ile                                                                                             160  - #               165  - #               170             - - Lys Gln Gly Pro Pro Arg Ser Lys Asp Glu Gl - #u Gln Lys Pro Gln Gln                  175      - #           180      - #           185                  - - Arg Pro Asp Leu Ala Val Asp Val Leu Glu Ar - #g Thr Ala Asp Lys Ala              190          - #       195          - #       200                      - - Thr Val Asn Gly Leu Pro Glu Lys Asp Arg Gl - #u Thr Asp Thr Ser Ala          205              - #   210              - #   215                          - - Leu Ala Ala Gly Ser Ser Gln Glu Val Thr Ty - #r Ala Gln Leu Asp His      220                 2 - #25                 2 - #30                 2 -      #35                                                                              - - Trp Ala Leu Thr Gln Arg Thr Ala Arg Ala Va - #l Ser Pro Gln Ser        Thr                                                                                             240  - #               245  - #               250             - - Lys Pro Met Ala Glu Ser Ile Thr Tyr Ala Al - #a Val Ala Arg His                      255      - #           260      - #           265                  - -  - - (2) INFORMATION FOR SEQ ID NO:9:                                     - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 568 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 24..428                                                - -     (ix) FEATURE:                                                                  (A) NAME/KEY: mat.sub.-- - #peptide                                           (B) LOCATION: 87..428                                                - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:9:                               - - CCACGCGTCC GGGGACCGGG GCC ATG TCT CCA CAC CTC AC - #T GCT CTC CTG             50                                                                                         - #        Met Ser Pro His Leu Thr - #Ala Leu Leu                             - #        -21 -20        - #         -15                    - - GGC CTA GTG CTC TGC CTG GCC CAG ACC ATC CA - #C ACG CAG GAG GGG GCC           98                                                                       Gly Leu Val Leu Cys Leu Ala Gln Thr Ile Hi - #s Thr Gln Glu Gly Ala                   -10          - #        -5          - #         1                      - - CTT CCC AGA CCC TCC ATC TCG GCT GAG CCA GG - #C ACT GTG ATC TCC CCG          146                                                                       Leu Pro Arg Pro Ser Ile Ser Ala Glu Pro Gl - #y Thr Val Ile Ser Pro             5                - #  10                - #  15                - #  20       - - GGG AGC CAT GTG ACT TTC ATG TGC CGG GGC CC - #G GTT GGG GTT CAA ACA          194                                                                       Gly Ser His Val Thr Phe Met Cys Arg Gly Pr - #o Val Gly Val Gln Thr                            25 - #                 30 - #                 35              - - TTC CGC CTG GAG AGG GAG GAT AGA GCC AAG TA - #C AAA GAT AGT TAT AAT          242                                                                       Phe Arg Leu Glu Arg Glu Asp Arg Ala Lys Ty - #r Lys Asp Ser Tyr Asn                        40     - #             45     - #             50                  - - GTG TTT CGA CTT GGT CCA TCT GAG TCA GAG GC - #C AGA TTC CAC ATT GAC          290                                                                       Val Phe Arg Leu Gly Pro Ser Glu Ser Glu Al - #a Arg Phe His Ile Asp                    55         - #         60         - #         65                      - - TCA GTA AGT GAA GGA AAT GCC GGG CTT TAT CG - #C TGC CTC TAT TAT AAG          338                                                                       Ser Val Ser Glu Gly Asn Ala Gly Leu Tyr Ar - #g Cys Leu Tyr Tyr Lys                70             - #     75             - #     80                          - - CCC CCT GGA TGG TCT GAG CAC AGT GAC TTC CT - #G GAG CTG CTG GTG AAA          386                                                                       Pro Pro Gly Trp Ser Glu His Ser Asp Phe Le - #u Glu Leu Leu Val Lys            85                 - # 90                 - # 95                 - #100       - - GGG ACT GTG CCA GGC ACT GAA GCC TCC GGA TT - #T GAT GCA CCA                 - # 428                                                                    Gly Thr Val Pro Gly Thr Glu Ala Ser Gly Ph - #e Asp Ala Pro                                   105  - #               110                                     - - TGAATGAGGA GAAATGGCCT CCCGTCTTGT GAACTTCAAT GGGGAGAAAT AA -             #TTAGAATG    488                                                                 - - AGCAATAGAA ATGCACAGAT GCCTATACAT ACATATACAA ATAAAAAGAT AC -            #GATTCGCA    548                                                                 - - AAAAAAAAAA AAAAAAGGGC            - #                  - #                      - #568                                                                  - -  - - (2) INFORMATION FOR SEQ ID NO:10:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 135 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:10:                              - - Met Ser Pro His Leu Thr Ala Leu Leu Gly Le - #u Val Leu Cys Leu Ala     21 -20                - # -15                - # -10                            - - Gln Thr Ile His Thr Gln Glu Gly Ala Leu Pr - #o Arg Pro Ser Ile Ser       -5                 - #  1               5 - #                 10              - - Ala Glu Pro Gly Thr Val Ile Ser Pro Gly Se - #r His Val Thr Phe Met                   15     - #             20     - #             25                  - - Cys Arg Gly Pro Val Gly Val Gln Thr Phe Ar - #g Leu Glu Arg Glu Asp               30         - #         35         - #         40                      - - Arg Ala Lys Tyr Lys Asp Ser Tyr Asn Val Ph - #e Arg Leu Gly Pro Ser           45             - #     50             - #     55                          - - Glu Ser Glu Ala Arg Phe His Ile Asp Ser Va - #l Ser Glu Gly Asn Ala       60                 - # 65                 - # 70                 - # 75       - - Gly Leu Tyr Arg Cys Leu Tyr Tyr Lys Pro Pr - #o Gly Trp Ser Glu His                       80 - #                 85 - #                 90              - - Ser Asp Phe Leu Glu Leu Leu Val Lys Gly Th - #r Val Pro Gly Thr Glu                   95     - #            100     - #            105                  - - Ala Ser Gly Phe Asp Ala Pro                                                      110                                                                    - -  - - (2) INFORMATION FOR SEQ ID NO:11:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 1620 base - #pairs                                                (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 81..1397                                               - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:11:                              - - GTCGACCCAC GCGTCCGCCT CTGTCCTGCC AGCACCGAGG GCTCATCCAT CC -             #ACAGAGCA     60                                                                 - - GTGCAGTGGG AGGAGACGCC ATG ACC CCC ATC CTC ACG GT - #C CTG ATC TGT            110                                                                                        - #    Met Thr Pro Ile Leu Thr Val Leu - #Ile Cys                             - #      1            - #   5               - #   10         - - CTC GGG CTG AGC CTG GAC CCC AGG ACC CAC GT - #G CAG GCA GGG CCC CTC          158                                                                       Leu Gly Leu Ser Leu Asp Pro Arg Thr His Va - #l Gln Ala Gly Pro Leu                            15 - #                 20 - #                 25              - - CCC AAG CCC ACC CTC TGG GCT GAG CCA GGC TC - #T GTG ATC ACC CAA GGG          206                                                                       Pro Lys Pro Thr Leu Trp Ala Glu Pro Gly Se - #r Val Ile Thr Gln Gly                        30     - #             35     - #             40                  - - AGT CCT GTG ACC CTC AGG TGT CAG GGG AGC CT - #G GAG ACG CAG GAG TAC          254                                                                       Ser Pro Val Thr Leu Arg Cys Gln Gly Ser Le - #u Glu Thr Gln Glu Tyr                    45         - #         50         - #         55                      - - CAT CTA TAT AGA GAA AAG AAA ACA GCA CTC TG - #G ATT ACA CGG ATC CCA          302                                                                       His Leu Tyr Arg Glu Lys Lys Thr Ala Leu Tr - #p Ile Thr Arg Ile Pro                60             - #     65             - #     70                          - - CAG GAG CTT GTG AAG AAG GGC CAG TTC CCC AT - #C CTA TCC ATC ACC TGG          350                                                                       Gln Glu Leu Val Lys Lys Gly Gln Phe Pro Il - #e Leu Ser Ile Thr Trp            75                 - # 80                 - # 85                 - # 90       - - GAA CAT GCA GGG CGG TAT TGC TGT ATC TAT GG - #C AGC CAC ACT GCA GGC          398                                                                       Glu His Ala Gly Arg Tyr Cys Cys Ile Tyr Gl - #y Ser His Thr Ala Gly                            95 - #                100 - #                105              - - CTC TCA GAG AGC AGT GAC CCC CTG GAG CTG GT - #G GTG ACA GGA GCC TAC          446                                                                       Leu Ser Glu Ser Ser Asp Pro Leu Glu Leu Va - #l Val Thr Gly Ala Tyr                       110      - #           115      - #           120                  - - AGC AAA CCC ACC CTC TCA GCT CTG CCC AGC CC - #T GTG GTG ACC TCA GGA          494                                                                       Ser Lys Pro Thr Leu Ser Ala Leu Pro Ser Pr - #o Val Val Thr Ser Gly                   125          - #       130          - #       135                      - - GGG AAT GTG ACC ATC CAG TGT GAC TCA CAG GT - #G GCA TTT GAT GGC TTC          542                                                                       Gly Asn Val Thr Ile Gln Cys Asp Ser Gln Va - #l Ala Phe Asp Gly Phe               140              - #   145              - #   150                          - - ATT CTG TGT AAG GAA GGA GAA GAT GAA CAC CC - #A CAA TGC CTG AAC TCC          590                                                                       Ile Leu Cys Lys Glu Gly Glu Asp Glu His Pr - #o Gln Cys Leu Asn Ser           155                 1 - #60                 1 - #65                 1 -      #70                                                                              - - CAT TCC CAT GCC CGT GGG TCA TCC CGG GCC AT - #C TTC TCC GTG GGC        CCC      638                                                                    His Ser His Ala Arg Gly Ser Ser Arg Ala Il - #e Phe Ser Val Gly Pro                          175  - #               180  - #               185              - - GTG AGC CCA AGT CGC AGG TGG TCG TAC AGG TG - #C TAT GGT TAT GAC TCG          686                                                                       Val Ser Pro Ser Arg Arg Trp Ser Tyr Arg Cy - #s Tyr Gly Tyr Asp Ser                       190      - #           195      - #           200                  - - CGC GCT CCC TAT GTG TGG TCT CTA CCC AGT GA - #T CTC CTG GGG CTC CTG          734                                                                       Arg Ala Pro Tyr Val Trp Ser Leu Pro Ser As - #p Leu Leu Gly Leu Leu                   205          - #       210          - #       215                      - - GTC CCA GGT GTT TCT AAG AAG CCA TCA CTC TC - #A GTG CAG CCG GGT CCT          782                                                                       Val Pro Gly Val Ser Lys Lys Pro Ser Leu Se - #r Val Gln Pro Gly Pro               220              - #   225              - #   230                          - - GTC GTG GCC CCT GGG GAG AAG CTG ACC TTC CA - #G TGT GGC TCT GAT GCC          830                                                                       Val Val Ala Pro Gly Glu Lys Leu Thr Phe Gl - #n Cys Gly Ser Asp Ala           235                 2 - #40                 2 - #45                 2 -      #50                                                                              - - GGC TAC GAC AGA TTT GTT CTG TAC AAG GAG TG - #G GGA CGT GAC TTC        CTC      878                                                                    Gly Tyr Asp Arg Phe Val Leu Tyr Lys Glu Tr - #p Gly Arg Asp Phe Leu                          255  - #               260  - #               265              - - CAG CGC CCT GGC CGG CAG CCC CAG GCT GGG CT - #C TCC CAG GCC AAC TTC          926                                                                       Gln Arg Pro Gly Arg Gln Pro Gln Ala Gly Le - #u Ser Gln Ala Asn Phe                       270      - #           275      - #           280                  - - ACC CTG GGC CCT GTG AGC CGC TCC TAC GGG GG - #C CAG TAC ACA TGC TCC          974                                                                       Thr Leu Gly Pro Val Ser Arg Ser Tyr Gly Gl - #y Gln Tyr Thr Cys Ser                   285          - #       290          - #       295                      - - GGT GCA TAC AAC CTC TCC TCC GAG TGG TCG GC - #C CCC AGC GAC CCC CTG         1022                                                                       Gly Ala Tyr Asn Leu Ser Ser Glu Trp Ser Al - #a Pro Ser Asp Pro Leu               300              - #   305              - #   310                          - - GAC ATC CTG ATC ACA GGA CAG ATC CGT GCC AG - #A CCC TTC CTC TCC GTG         1070                                                                       Asp Ile Leu Ile Thr Gly Gln Ile Arg Ala Ar - #g Pro Phe Leu Ser Val           315                 3 - #20                 3 - #25                 3 -      #30                                                                              - - CGG CCG GGC CCC ACA GTG GCC TCA GGA GAG AA - #C GTG ACC CTG CTG        TGT     1118                                                                    Arg Pro Gly Pro Thr Val Ala Ser Gly Glu As - #n Val Thr Leu Leu Cys                          335  - #               340  - #               345              - - CAG TCA CAG GGA GGG ATG CAC ACT TTC CTT TT - #G ACC AAG GAG GGG GCA         1166                                                                       Gln Ser Gln Gly Gly Met His Thr Phe Leu Le - #u Thr Lys Glu Gly Ala                       350      - #           355      - #           360                  - - GCT GAT TCC CCG CTG CGT CTA AAA TCA AAG CG - #C CAA TCT CAT AAG TAC         1214                                                                       Ala Asp Ser Pro Leu Arg Leu Lys Ser Lys Ar - #g Gln Ser His Lys Tyr                   365          - #       370          - #       375                      - - CAG GCT GAA TTC CCC ATG AGT CCT GTG ACC TC - #G GCC CAC GCG GGG ACC         1262                                                                       Gln Ala Glu Phe Pro Met Ser Pro Val Thr Se - #r Ala His Ala Gly Thr               380              - #   385              - #   390                          - - TAC AGG TGC TAC GGC TCA CTC AGC TCC AAC CC - #C TAC CTG CTG ACT CAC         1310                                                                       Tyr Arg Cys Tyr Gly Ser Leu Ser Ser Asn Pr - #o Tyr Leu Leu Thr His           395                 4 - #00                 4 - #05                 4 -      #10                                                                              - - CCC AGT GAC CCC CTG GAG CTC GTG GTC TCA GG - #A GCA GCT GAG ACC        CTC     1358                                                                    Pro Ser Asp Pro Leu Glu Leu Val Val Ser Gl - #y Ala Ala Glu Thr Leu                          415  - #               420  - #               425              - - AGC CCA CCA CAA AAC AAG TCC GAC TCC AAG GC - #T GGT GAG TGAGGAGATG          1407                                                                       Ser Pro Pro Gln Asn Lys Ser Asp Ser Lys Al - #a Gly Glu                                   430      - #           435                                         - - CTTGCCGTGA TGACGCTGGG CACAGAGGGT CAGGTCCTGT CAAGAGGAGC TG -             #GGTGTCCT   1467                                                                 - - GGGTGGACAT TTGAAGAATT ATATTCATTC CAACTTGAAG AATTATTCAA CA -            #CCTTTAAC   1527                                                                 - - AATGTATATG TGAAGTACTT TATTCTTTCA TATTTTAAAA ATAAAAGATA AT -            #TATCCATG   1587                                                                 - - AAAAAAAAAA AAAAAAAAAA AAAGGGCGGC CGC       - #                  -      #       1620                                                                     - -  - - (2) INFORMATION FOR SEQ ID NO:12:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 439 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:12:                              - - Met Thr Pro Ile Leu Thr Val Leu Ile Cys Le - #u Gly Leu Ser Leu        Asp                                                                               1               5 - #                 10 - #                 15             - - Pro Arg Thr His Val Gln Ala Gly Pro Leu Pr - #o Lys Pro Thr Leu Trp                   20     - #             25     - #             30                  - - Ala Glu Pro Gly Ser Val Ile Thr Gln Gly Se - #r Pro Val Thr Leu Arg               35         - #         40         - #         45                      - - Cys Gln Gly Ser Leu Glu Thr Gln Glu Tyr Hi - #s Leu Tyr Arg Glu Lys           50             - #     55             - #     60                          - - Lys Thr Ala Leu Trp Ile Thr Arg Ile Pro Gl - #n Glu Leu Val Lys Lys       65                 - # 70                 - # 75                 - # 80       - - Gly Gln Phe Pro Ile Leu Ser Ile Thr Trp Gl - #u His Ala Gly Arg Tyr                       85 - #                 90 - #                 95              - - Cys Cys Ile Tyr Gly Ser His Thr Ala Gly Le - #u Ser Glu Ser Ser Asp                  100      - #           105      - #           110                  - - Pro Leu Glu Leu Val Val Thr Gly Ala Tyr Se - #r Lys Pro Thr Leu Ser              115          - #       120          - #       125                      - - Ala Leu Pro Ser Pro Val Val Thr Ser Gly Gl - #y Asn Val Thr Ile Gln          130              - #   135              - #   140                          - - Cys Asp Ser Gln Val Ala Phe Asp Gly Phe Il - #e Leu Cys Lys Glu Gly      145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - Glu Asp Glu His Pro Gln Cys Leu Asn Ser Hi - #s Ser His Ala Arg        Gly                                                                                             165  - #               170  - #               175             - - Ser Ser Arg Ala Ile Phe Ser Val Gly Pro Va - #l Ser Pro Ser Arg Arg                  180      - #           185      - #           190                  - - Trp Ser Tyr Arg Cys Tyr Gly Tyr Asp Ser Ar - #g Ala Pro Tyr Val Trp              195          - #       200          - #       205                      - - Ser Leu Pro Ser Asp Leu Leu Gly Leu Leu Va - #l Pro Gly Val Ser Lys          210              - #   215              - #   220                          - - Lys Pro Ser Leu Ser Val Gln Pro Gly Pro Va - #l Val Ala Pro Gly Glu      225                 2 - #30                 2 - #35                 2 -      #40                                                                              - - Lys Leu Thr Phe Gln Cys Gly Ser Asp Ala Gl - #y Tyr Asp Arg Phe        Val                                                                                             245  - #               250  - #               255             - - Leu Tyr Lys Glu Trp Gly Arg Asp Phe Leu Gl - #n Arg Pro Gly Arg Gln                  260      - #           265      - #           270                  - - Pro Gln Ala Gly Leu Ser Gln Ala Asn Phe Th - #r Leu Gly Pro Val Ser              275          - #       280          - #       285                      - - Arg Ser Tyr Gly Gly Gln Tyr Thr Cys Ser Gl - #y Ala Tyr Asn Leu Ser          290              - #   295              - #   300                          - - Ser Glu Trp Ser Ala Pro Ser Asp Pro Leu As - #p Ile Leu Ile Thr Gly      305                 3 - #10                 3 - #15                 3 -      #20                                                                              - - Gln Ile Arg Ala Arg Pro Phe Leu Ser Val Ar - #g Pro Gly Pro Thr        Val                                                                                             325  - #               330  - #               335             - - Ala Ser Gly Glu Asn Val Thr Leu Leu Cys Gl - #n Ser Gln Gly Gly Met                  340      - #           345      - #           350                  - - His Thr Phe Leu Leu Thr Lys Glu Gly Ala Al - #a Asp Ser Pro Leu Arg              355          - #       360          - #       365                      - - Leu Lys Ser Lys Arg Gln Ser His Lys Tyr Gl - #n Ala Glu Phe Pro Met          370              - #   375              - #   380                          - - Ser Pro Val Thr Ser Ala His Ala Gly Thr Ty - #r Arg Cys Tyr Gly Ser      385                 3 - #90                 3 - #95                 4 -      #00                                                                              - - Leu Ser Ser Asn Pro Tyr Leu Leu Thr His Pr - #o Ser Asp Pro Leu        Glu                                                                                             405  - #               410  - #               415             - - Leu Val Val Ser Gly Ala Ala Glu Thr Leu Se - #r Pro Pro Gln Asn Lys                  420      - #           425      - #           430                  - - Ser Asp Ser Lys Ala Gly Glu                                                      435                                                                    - -  - - (2) INFORMATION FOR SEQ ID NO:13:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 2197 base - #pairs                                                (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 191..1483                                              - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:13:                              - - GTCGACCCAC GCGTCCGGTC AACTTTTCTT CCCCTACTTC CCTGCATTTC TC -             #CTCTGTGC     60                                                                 - - TCACTGCCAC ACGCAGCTCA ACCTGGACGG CACAGCCAGA TGCGAGATGC GT -            #CTCTGCTG    120                                                                 - - ATCTGAGTCT GCCTGCAGCA TGGACCTGGG TCTTCCCTGA AGCATCTCCA GG -            #GCTGGAGG    180                                                                 - - GACGACTGCC ATG CAC CGA GGG CTC ATC CAT CCG CA - #G AGC AGG GCA GTG           229                                                                                 Met His Arg Gly - #Leu Ile His Pro Gln Ser Arg Ala Val                          1     - #          5        - #          10                       - - GGA GGA GAC GCC ATG ACC CCC ATC GTC ACA GT - #C CTG ATC TGT CTC GGG          277                                                                       Gly Gly Asp Ala Met Thr Pro Ile Val Thr Va - #l Leu Ile Cys Leu Gly                15             - #     20             - #     25                          - - CTG AGT CTG GGC CCC AGG ACC CAC GTG CAG AC - #A GGG ACC ATC CCC AAG          325                                                                       Leu Ser Leu Gly Pro Arg Thr His Val Gln Th - #r Gly Thr Ile Pro Lys            30                 - # 35                 - # 40                 - # 45       - - CCC ACC CTG TGG GCT GAG CCA GAC TCT GTG AT - #C ACC CAG GGG AGT CCC          373                                                                       Pro Thr Leu Trp Ala Glu Pro Asp Ser Val Il - #e Thr Gln Gly Ser Pro                            50 - #                 55 - #                 60              - - GTC ACC CTC AGT TGT CAG GGG AGC CTT GAA GC - #C CAG GAG TAC CGT CTA          421                                                                       Val Thr Leu Ser Cys Gln Gly Ser Leu Glu Al - #a Gln Glu Tyr Arg Leu                        65     - #             70     - #             75                  - - TAT AGG GAG AAA AAA TCA GCA TCT TGG ATT AC - #A CGG ATA CGA CCA GAG          469                                                                       Tyr Arg Glu Lys Lys Ser Ala Ser Trp Ile Th - #r Arg Ile Arg Pro Glu                    80         - #         85         - #         90                      - - CTT GTG AAG AAC GGC CAG TTC CAC ATC CCA TC - #C ATC ACC TGG GAA CAC          517                                                                       Leu Val Lys Asn Gly Gln Phe His Ile Pro Se - #r Ile Thr Trp Glu His                95             - #    100             - #    105                          - - ACA GGG CGA TAT GGC TGT CAG TAT TAC AGC CG - #C GCT CGG TGG TCT GAG          565                                                                       Thr Gly Arg Tyr Gly Cys Gln Tyr Tyr Ser Ar - #g Ala Arg Trp Ser Glu           110                 1 - #15                 1 - #20                 1 -      #25                                                                              - - CTC AGT GAC CCC CTG GTG CTG GTG ATG ACA GG - #A GCC TAC CCA AAA        CCC      613                                                                    Leu Ser Asp Pro Leu Val Leu Val Met Thr Gl - #y Ala Tyr Pro Lys Pro                          130  - #               135  - #               140              - - ACC CTC TCA GCC CAG CCC AGC CCT GTG GTG AC - #C TCA GGA GGA AGG GTG          661                                                                       Thr Leu Ser Ala Gln Pro Ser Pro Val Val Th - #r Ser Gly Gly Arg Val                       145      - #           150      - #           155                  - - ACC CTC CAG TGT GAG TCA CAG GTG GCA TTT GG - #C GGC TTC ATT CTG TGT          709                                                                       Thr Leu Gln Cys Glu Ser Gln Val Ala Phe Gl - #y Gly Phe Ile Leu Cys                   160          - #       165          - #       170                      - - AAG GAA GGA GAA GAT GAA CAC CCA CAA TGC CT - #G AAC TCC CAG CCC CAT          757                                                                       Lys Glu Gly Glu Asp Glu His Pro Gln Cys Le - #u Asn Ser Gln Pro His               175              - #   180              - #   185                          - - GCC CGT GGG TCG TCC CGC GCC ATC TTC TCC GT - #G GGC CCC GTG AGC CCG          805                                                                       Ala Arg Gly Ser Ser Arg Ala Ile Phe Ser Va - #l Gly Pro Val Ser Pro           190                 1 - #95                 2 - #00                 2 -      #05                                                                              - - AAT CGC AGG TGG TCG CAC AGG TGC TAT GGT TA - #T GAC TTG AAC TCT        CCC      853                                                                    Asn Arg Arg Trp Ser His Arg Cys Tyr Gly Ty - #r Asp Leu Asn Ser Pro                          210  - #               215  - #               220              - - TAT GTG TGG TCT TCA CCC AGT GAT CTC CTG GA - #G CTC CTG GTC CCA GGT          901                                                                       Tyr Val Trp Ser Ser Pro Ser Asp Leu Leu Gl - #u Leu Leu Val Pro Gly                       225      - #           230      - #           235                  - - GTT TCT AAG AAG CCA TCA CTC TCA GTG CAG CC - #G GGT CCT GTC GTG GCC          949                                                                       Val Ser Lys Lys Pro Ser Leu Ser Val Gln Pr - #o Gly Pro Val Val Ala                   240          - #       245          - #       250                      - - CCT GGG GAA AGC CTG ACC CTC CAG TGT GTC TC - #T GAT GTC GGC TAT GAC          997                                                                       Pro Gly Glu Ser Leu Thr Leu Gln Cys Val Se - #r Asp Val Gly Tyr Asp               255              - #   260              - #   265                          - - AGA TTT GTT CTG TAC AAG GAG GGG GAA CGT GA - #C CTT CGC CAG CTC CCT         1045                                                                       Arg Phe Val Leu Tyr Lys Glu Gly Glu Arg As - #p Leu Arg Gln Leu Pro           270                 2 - #75                 2 - #80                 2 -      #85                                                                              - - GGC CGG CAG CCC CAG GCT GGG CTC TCC CAG GC - #C AAC TTC ACC CTG        GGC     1093                                                                    Gly Arg Gln Pro Gln Ala Gly Leu Ser Gln Al - #a Asn Phe Thr Leu Gly                          290  - #               295  - #               300              - - CCT GTG AGC CGC TCC TAC GGG GGC CAG TAC AG - #A TGC TAC GGT GCA TAC         1141                                                                       Pro Val Ser Arg Ser Tyr Gly Gly Gln Tyr Ar - #g Cys Tyr Gly Ala Tyr                       305      - #           310      - #           315                  - - AAC CTC TCC TCC GAG TGG TCG GCC CCC AGC GA - #C CCC CTG GAC ATC CTG         1189                                                                       Asn Leu Ser Ser Glu Trp Ser Ala Pro Ser As - #p Pro Leu Asp Ile Leu                   320          - #       325          - #       330                      - - ATC ACA GGA CAG ATC CAT GGC ACA CCC TTC AT - #C TCA GTG CAG CCA GGC         1237                                                                       Ile Thr Gly Gln Ile His Gly Thr Pro Phe Il - #e Ser Val Gln Pro Gly               335              - #   340              - #   345                          - - CCC ACA GTG GCC TCA GGA GAG AAC GTG ACC CT - #G CTG TGT CAG TCA TGG         1285                                                                       Pro Thr Val Ala Ser Gly Glu Asn Val Thr Le - #u Leu Cys Gln Ser Trp           350                 3 - #55                 3 - #60                 3 -      #65                                                                              - - CGG CAG TTC CAC ACT TTC CTT CTG ACC AAG GC - #G GGA GCA GCT GAT        GCC     1333                                                                    Arg Gln Phe His Thr Phe Leu Leu Thr Lys Al - #a Gly Ala Ala Asp Ala                          370  - #               375  - #               380              - - CCA CTC CGT CTA AGA TCA ATA CAC GAA TAT CC - #T AAG TAC CAG GCT GAA         1381                                                                       Pro Leu Arg Leu Arg Ser Ile His Glu Tyr Pr - #o Lys Tyr Gln Ala Glu                       385      - #           390      - #           395                  - - TTC CCC ATG AGT CCT GTG ACC TCA GCC CAC GC - #G GGG ACC TAC AGG ACC         1429                                                                       Phe Pro Met Ser Pro Val Thr Ser Ala His Al - #a Gly Thr Tyr Arg Thr                   400          - #       405          - #       410                      - - CTC CAT GGG TTC CAG CCC CCC ACC CAC CGG TC - #C CAT CTC CAC ACC TGC         1477                                                                       Leu His Gly Phe Gln Pro Pro Thr His Arg Se - #r His Leu His Thr Cys               415              - #   420              - #   425                          - - AGG CCC TGAGGACCAG CCCCTCACCC CCACTGGGTC GGATCCCCAA AG - #TGGTCTGG          1533                                                                       Arg Pro                                                                       430                                                                            - - GAAGGCACCT GGGGGTTGTG ATCGGCATCT TGGTGGCCGT CGTCCTACTG CT -             #CCTCCTCC   1593                                                                 - - TCCTCCTCCT CTTCCTCATC CTCCGACATC GACGTCAGGG CAAACACTGG AC -            #ATCGACCC   1653                                                                 - - AGAGAAAGGC TGATTTCCAA CATCCTGCAG GGGCTGTGGG GCCAGAGCCC AC -            #AGACAGAG   1713                                                                 - - GCCTGCAGTG GAGGTCCAGC CCAGCTGCCG ACGCCCAGGA AGAAAACCTC TA -            #TGCTGCCG   1773                                                                 - - TGAAGGACAC ACAGCCTGAA GATGGGGTGG AGATGGACAC TCGGGCTGCT GC -            #ATCTGAAG   1833                                                                 - - CCCCCCAGGA TGTGACCTAC GCCCAGCTGC ACAGCTTGAC CCTCAGACGG AA -            #GGCAACTG   1893                                                                 - - AGCCTCCTCC ATCCCAGGAA AGGGAACCTC CAGCTGAGCC CAGCATTTAC GC -            #CACCCTGG   1953                                                                 - - CCATCCACTA GCCCGGAGGG TACGCAGACT CCACACTCAG TAGAAGGAGA CT -            #CAGGACTG   2013                                                                 - - CTGAAGGCAC GGGAGCTGCC CCCAGTGGAC ACCAATGAAC CCCAGTCAGC CT -            #GGACCCCT   2073                                                                 - - AACAAAGACC ATGAGGAGAT GCTGGGAACT TTGGGACTCA CTTGATTCTG CA -            #GTGGAAAT   2133                                                                 - - AACTAATATC CCTACATTTT TTAATTAAAG CAACAGACTT CTCAATAATC AA -            #TGAGTTAA   2193                                                                 - - CCGA                 - #                  - #                  - #               2197                                                                  - -  - - (2) INFORMATION FOR SEQ ID NO:14:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 431 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:14:                              - - Met His Arg Gly Leu Ile His Pro Gln Ser Ar - #g Ala Val Gly Gly Asp        1               5 - #                 10 - #                 15              - - Ala Met Thr Pro Ile Val Thr Val Leu Ile Cy - #s Leu Gly Leu Ser Leu                   20     - #             25     - #             30                  - - Gly Pro Arg Thr His Val Gln Thr Gly Thr Il - #e Pro Lys Pro Thr Leu               35         - #         40         - #         45                      - - Trp Ala Glu Pro Asp Ser Val Ile Thr Gln Gl - #y Ser Pro Val Thr Leu           50             - #     55             - #     60                          - - Ser Cys Gln Gly Ser Leu Glu Ala Gln Glu Ty - #r Arg Leu Tyr Arg Glu       65                 - # 70                 - # 75                 - # 80       - - Lys Lys Ser Ala Ser Trp Ile Thr Arg Ile Ar - #g Pro Glu Leu Val Lys                       85 - #                 90 - #                 95              - - Asn Gly Gln Phe His Ile Pro Ser Ile Thr Tr - #p Glu His Thr Gly Arg                  100      - #           105      - #           110                  - - Tyr Gly Cys Gln Tyr Tyr Ser Arg Ala Arg Tr - #p Ser Glu Leu Ser Asp              115          - #       120          - #       125                      - - Pro Leu Val Leu Val Met Thr Gly Ala Tyr Pr - #o Lys Pro Thr Leu Ser          130              - #   135              - #   140                          - - Ala Gln Pro Ser Pro Val Val Thr Ser Gly Gl - #y Arg Val Thr Leu Gln      145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - Cys Glu Ser Gln Val Ala Phe Gly Gly Phe Il - #e Leu Cys Lys Glu        Gly                                                                                             165  - #               170  - #               175             - - Glu Asp Glu His Pro Gln Cys Leu Asn Ser Gl - #n Pro His Ala Arg Gly                  180      - #           185      - #           190                  - - Ser Ser Arg Ala Ile Phe Ser Val Gly Pro Va - #l Ser Pro Asn Arg Arg              195          - #       200          - #       205                      - - Trp Ser His Arg Cys Tyr Gly Tyr Asp Leu As - #n Ser Pro Tyr Val Trp          210              - #   215              - #   220                          - - Ser Ser Pro Ser Asp Leu Leu Glu Leu Leu Va - #l Pro Gly Val Ser Lys      225                 2 - #30                 2 - #35                 2 -      #40                                                                              - - Lys Pro Ser Leu Ser Val Gln Pro Gly Pro Va - #l Val Ala Pro Gly        Glu                                                                                             245  - #               250  - #               255             - - Ser Leu Thr Leu Gln Cys Val Ser Asp Val Gl - #y Tyr Asp Arg Phe Val                  260      - #           265      - #           270                  - - Leu Tyr Lys Glu Gly Glu Arg Asp Leu Arg Gl - #n Leu Pro Gly Arg Gln              275          - #       280          - #       285                      - - Pro Gln Ala Gly Leu Ser Gln Ala Asn Phe Th - #r Leu Gly Pro Val Ser          290              - #   295              - #   300                          - - Arg Ser Tyr Gly Gly Gln Tyr Arg Cys Tyr Gl - #y Ala Tyr Asn Leu Ser      305                 3 - #10                 3 - #15                 3 -      #20                                                                              - - Ser Glu Trp Ser Ala Pro Ser Asp Pro Leu As - #p Ile Leu Ile Thr        Gly                                                                                             325  - #               330  - #               335             - - Gln Ile His Gly Thr Pro Phe Ile Ser Val Gl - #n Pro Gly Pro Thr Val                  340      - #           345      - #           350                  - - Ala Ser Gly Glu Asn Val Thr Leu Leu Cys Gl - #n Ser Trp Arg Gln Phe              355          - #       360          - #       365                      - - His Thr Phe Leu Leu Thr Lys Ala Gly Ala Al - #a Asp Ala Pro Leu Arg          370              - #   375              - #   380                          - - Leu Arg Ser Ile His Glu Tyr Pro Lys Tyr Gl - #n Ala Glu Phe Pro Met      385                 3 - #90                 3 - #95                 4 -      #00                                                                              - - Ser Pro Val Thr Ser Ala His Ala Gly Thr Ty - #r Arg Thr Leu His        Gly                                                                                             405  - #               410  - #               415             - - Phe Gln Pro Pro Thr His Arg Ser His Leu Hi - #s Thr Cys Arg Pro                      420      - #           425      - #           430                  - -  - - (2) INFORMATION FOR SEQ ID NO:15:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 2271 base - #pairs                                                (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 191..2035                                              - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:15:                              - - GTCGACCCAC GCGTCCGGTC AACTTTTCTT CCCCTACTTC CCTGCATTTC TC -             #CTCTGTGC     60                                                                 - - TCACTGCCAC ACGCAGCTCA ACCTGGACGG CACAGCCAGA TGCGAGATGC GT -            #CTCTGCTG    120                                                                 - - ATCTGAGTCT GCCTGCAGCA TGGACCTGGG TCTTCCCTGA AGCATCTCCA GG -            #GCTGGAGG    180                                                                 - - GACGACTGCC ATG CAC CGA GGG CTC ATC CAT CCG CA - #G AGC AGG GCA GTG           229                                                                                 Met His Arg Gly - #Leu Ile His Pro Gln Ser Arg Ala Val                          1     - #          5        - #          10                       - - GGA GGA GAC GCC ATG ACC CCC ATC GTC ACA GT - #C CTG ATC TGT CTC GGG          277                                                                       Gly Gly Asp Ala Met Thr Pro Ile Val Thr Va - #l Leu Ile Cys Leu Gly                15             - #     20             - #     25                          - - CTG AGT CTG GGC CCC AGG ACC CAC GTG CAG AC - #A GGG ACC ATC CCC AAG          325                                                                       Leu Ser Leu Gly Pro Arg Thr His Val Gln Th - #r Gly Thr Ile Pro Lys            30                 - # 35                 - # 40                 - # 45       - - CCC ACC CTG TGG GCT GAG CCA GAC TCT GTG AT - #C ACC CAG GGG AGT CCC          373                                                                       Pro Thr Leu Trp Ala Glu Pro Asp Ser Val Il - #e Thr Gln Gly Ser Pro                            50 - #                 55 - #                 60              - - GTC ACC CTC AGT TGT CAG GGG AGC CTT GAA GC - #C CAG GAG TAC CGT CTA          421                                                                       Val Thr Leu Ser Cys Gln Gly Ser Leu Glu Al - #a Gln Glu Tyr Arg Leu                        65     - #             70     - #             75                  - - TAT AGG GAG AAA AAA TCA GCA TCT TGG ATT AC - #A CGG ATA CGA CCA GAG          469                                                                       Tyr Arg Glu Lys Lys Ser Ala Ser Trp Ile Th - #r Arg Ile Arg Pro Glu                    80         - #         85         - #         90                      - - CTT GTG AAG AAC GGC CAG TTC CAC ATC CCA TC - #C ATC ACC TGG GAA CAC          517                                                                       Leu Val Lys Asn Gly Gln Phe His Ile Pro Se - #r Ile Thr Trp Glu His                95             - #    100             - #    105                          - - ACA GGG CGA TAT GGC TGT CAG TAT TAC AGC CG - #C GCT CGG TGG TCT GAG          565                                                                       Thr Gly Arg Tyr Gly Cys Gln Tyr Tyr Ser Ar - #g Ala Arg Trp Ser Glu           110                 1 - #15                 1 - #20                 1 -      #25                                                                              - - CTC AGT GAC CCC CTG GTG CTG GTG ATG ACA GG - #A GCC TAC CCA AAA        CCC      613                                                                    Leu Ser Asp Pro Leu Val Leu Val Met Thr Gl - #y Ala Tyr Pro Lys Pro                          130  - #               135  - #               140              - - ACC CTC TCA GCC CAG CCC AGC CCT GTG GTG AC - #C TCA GGA GGA AGG GTG          661                                                                       Thr Leu Ser Ala Gln Pro Ser Pro Val Val Th - #r Ser Gly Gly Arg Val                       145      - #           150      - #           155                  - - ACC CTC CAG TGT GAG TCA CAG GTG GCA TTT GG - #C GGC TTC ATT CTG TGT          709                                                                       Thr Leu Gln Cys Glu Ser Gln Val Ala Phe Gl - #y Gly Phe Ile Leu Cys                   160          - #       165          - #       170                      - - AAG GAA GGA GAA GAT GAA CAC CCA CAA TGC CT - #G AAC TCC CAG CCC CAT          757                                                                       Lys Glu Gly Glu Asp Glu His Pro Gln Cys Le - #u Asn Ser Gln Pro His               175              - #   180              - #   185                          - - GCC CGT GGG TCG TCC CGC GCC ATC TTC TCC GT - #G GGC CCC GTG AGC CCG          805                                                                       Ala Arg Gly Ser Ser Arg Ala Ile Phe Ser Va - #l Gly Pro Val Ser Pro           190                 1 - #95                 2 - #00                 2 -      #05                                                                              - - AAT CGC AGG TGG TCG CAC AGG TGC TAT GGT TA - #T GAC TTG AAC TCT        CCC      853                                                                    Asn Arg Arg Trp Ser His Arg Cys Tyr Gly Ty - #r Asp Leu Asn Ser Pro                          210  - #               215  - #               220              - - TAT GTG TGG TCT TCA CCC AGT GAT CTC CTG GA - #G CTC CTG GTC CCA GGT          901                                                                       Tyr Val Trp Ser Ser Pro Ser Asp Leu Leu Gl - #u Leu Leu Val Pro Gly                       225      - #           230      - #           235                  - - GTT TCT AAG AAG CCA TCA CTC TCA GTG CAG CC - #G GGT CCT GTC GTG GCC          949                                                                       Val Ser Lys Lys Pro Ser Leu Ser Val Gln Pr - #o Gly Pro Val Val Ala                   240          - #       245          - #       250                      - - CCT GGG GAA AGC CTG ACC CTC CAG TGT GTC TC - #T GAT GTC GGC TAT GAC          997                                                                       Pro Gly Glu Ser Leu Thr Leu Gln Cys Val Se - #r Asp Val Gly Tyr Asp               255              - #   260              - #   265                          - - AGA TTT GTT CTG TAC AAG GAG GGG GAA CGT GA - #C CTT CGC CAG CTC CCT         1045                                                                       Arg Phe Val Leu Tyr Lys Glu Gly Glu Arg As - #p Leu Arg Gln Leu Pro           270                 2 - #75                 2 - #80                 2 -      #85                                                                              - - GGC CGG CAG CCC CAG GCT GGG CTC TCC CAG GC - #C AAC TTC ACC CTG        GGC     1093                                                                    Gly Arg Gln Pro Gln Ala Gly Leu Ser Gln Al - #a Asn Phe Thr Leu Gly                          290  - #               295  - #               300              - - CCT GTG AGC CGC TCC TAC GGG GGC CAG TAC AG - #A TGC TAC GGT GCA TAC         1141                                                                       Pro Val Ser Arg Ser Tyr Gly Gly Gln Tyr Ar - #g Cys Tyr Gly Ala Tyr                       305      - #           310      - #           315                  - - AAC CTC TCC TCC GAG TGG TCG GCC CCC AGC GA - #C CCC CTG GAC ATC CTG         1189                                                                       Asn Leu Ser Ser Glu Trp Ser Ala Pro Ser As - #p Pro Leu Asp Ile Leu                   320          - #       325          - #       330                      - - ATC ACA GGA CAG ATC CAT GGC ACA CCC TTC AT - #C TCA GTG CAG CCA GGC         1237                                                                       Ile Thr Gly Gln Ile His Gly Thr Pro Phe Il - #e Ser Val Gln Pro Gly               335              - #   340              - #   345                          - - CCC ACA GTG GCC TCA GGA GAG AAC GTG ACC CT - #G CTG TGT CAG TCA TGG         1285                                                                       Pro Thr Val Ala Ser Gly Glu Asn Val Thr Le - #u Leu Cys Gln Ser Trp           350                 3 - #55                 3 - #60                 3 -      #65                                                                              - - CGG CAG TTC CAC ACT TTC CTT CTG ACC AAG GC - #G GGA GCA GCT GAT        GCC     1333                                                                    Arg Gln Phe His Thr Phe Leu Leu Thr Lys Al - #a Gly Ala Ala Asp Ala                          370  - #               375  - #               380              - - CCA CTC CGT CTA AGA TCA ATA CAC GAA TAT CC - #T AAG TAC CAG GCT GAA         1381                                                                       Pro Leu Arg Leu Arg Ser Ile His Glu Tyr Pr - #o Lys Tyr Gln Ala Glu                       385      - #           390      - #           395                  - - TTC CCC ATG AGT CCT GTG ACC TCA GCC CAC GC - #G GGG ACC TAC AGG TGC         1429                                                                       Phe Pro Met Ser Pro Val Thr Ser Ala His Al - #a Gly Thr Tyr Arg Cys                   400          - #       405          - #       410                      - - TAC GGC TCA CTC AAC TCC GAC CCC TAC CTG CT - #G TCT CAC CCC AGT GAG         1477                                                                       Tyr Gly Ser Leu Asn Ser Asp Pro Tyr Leu Le - #u Ser His Pro Ser Glu               415              - #   420              - #   425                          - - CCC CTG GAG CTC GTG GTC TCA GGA CCC TCC AT - #G GGT TCC AGC CCC CCA         1525                                                                       Pro Leu Glu Leu Val Val Ser Gly Pro Ser Me - #t Gly Ser Ser Pro Pro           430                 4 - #35                 4 - #40                 4 -      #45                                                                              - - CCC ACC GGT CCC ATC TCC ACA CCT GCA GGC CC - #T GAG GAC CAG CCC        CTC     1573                                                                    Pro Thr Gly Pro Ile Ser Thr Pro Ala Gly Pr - #o Glu Asp Gln Pro Leu                          450  - #               455  - #               460              - - ACC CCC ACT GGG TCG GAT CCC CAA AGT GGT CT - #G GGA AGG CAC CTG GGG         1621                                                                       Thr Pro Thr Gly Ser Asp Pro Gln Ser Gly Le - #u Gly Arg His Leu Gly                       465      - #           470      - #           475                  - - GTT GTG ATC GGC ATC TTG GTG GCC GTC GTC CT - #A CTG CTC CTC CTC CTC         1669                                                                       Val Val Ile Gly Ile Leu Val Ala Val Val Le - #u Leu Leu Leu Leu Leu                   480          - #       485          - #       490                      - - CTC CTC CTC TTC CTC ATC CTC CGA CAT CGA CG - #T CAG GGC AAA CAC TGG         1717                                                                       Leu Leu Leu Phe Leu Ile Leu Arg His Arg Ar - #g Gln Gly Lys His Trp               495              - #   500              - #   505                          - - ACA TCG ACC CAG AGA AAG GCT GAT TTC CAA CA - #T CCT GCA GGG GCT GTG         1765                                                                       Thr Ser Thr Gln Arg Lys Ala Asp Phe Gln Hi - #s Pro Ala Gly Ala Val           510                 5 - #15                 5 - #20                 5 -      #25                                                                              - - GGG CCA GAG CCC ACA GAC AGA GGC CTG CAG TG - #G AGG TCC AGC CCA        GCT     1813                                                                    Gly Pro Glu Pro Thr Asp Arg Gly Leu Gln Tr - #p Arg Ser Ser Pro Ala                          530  - #               535  - #               540              - - GCC GAC GCC CAG GAA GAA AAC CTC TAT GCT GC - #C GTG AAG GAC ACA CAG         1861                                                                       Ala Asp Ala Gln Glu Glu Asn Leu Tyr Ala Al - #a Val Lys Asp Thr Gln                       545      - #           550      - #           555                  - - CCT GAA GAT GGG GTG GAG ATG GAC ACT CGG GC - #T GCT GCA TCT GAA GCC         1909                                                                       Pro Glu Asp Gly Val Glu Met Asp Thr Arg Al - #a Ala Ala Ser Glu Ala                   560          - #       565          - #       570                      - - CCC CAG GAT GTG ACC TAC GCC CAG CTG CAC AG - #C TTG ACC CTC AGA CGG         1957                                                                       Pro Gln Asp Val Thr Tyr Ala Gln Leu His Se - #r Leu Thr Leu Arg Arg               575              - #   580              - #   585                          - - AAG GCA ACT GAG CCT CCT CCA TCC CAG GAA AG - #G GAA CCT CCA GCT GAG         2005                                                                       Lys Ala Thr Glu Pro Pro Pro Ser Gln Glu Ar - #g Glu Pro Pro Ala Glu           590                 5 - #95                 6 - #00                 6 -      #05                                                                              - - CCC AGC ATT TAC GCC ACC CTG GCC ATC CAC TA - #GCCCGGAG GGTACGCAGA           2055                                                                      Pro Ser Ile Tyr Ala Thr Leu Ala Ile His                                                       610  - #               615                                     - - CTCCACACTC AGTAGAAGGA GACTCAGGAC TGCTGAAGGC ACGGGAGCTG CC -             #CCCAGTGG   2115                                                                 - - ACACCAATGA ACCCCAGTCA GCCTGGACCC CTAACAAAGA CCATGAGGAG AT -            #GCTGGGAA   2175                                                                 - - CTTTGGGACT CACTTGATTC TGCAGTGGAA ATAACTAATA TCCCTACATT TT -            #TTAATTAA   2235                                                                 - - AGCAACAGAC TTCTCAATAA TCAATGAGTT AACCGA      - #                       - #     2271                                                                     - -  - - (2) INFORMATION FOR SEQ ID NO:16:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 615 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:16:                              - - Met His Arg Gly Leu Ile His Pro Gln Ser Ar - #g Ala Val Gly Gly        Asp                                                                               1               5 - #                 10 - #                 15             - - Ala Met Thr Pro Ile Val Thr Val Leu Ile Cy - #s Leu Gly Leu Ser Leu                   20     - #             25     - #             30                  - - Gly Pro Arg Thr His Val Gln Thr Gly Thr Il - #e Pro Lys Pro Thr Leu               35         - #         40         - #         45                      - - Trp Ala Glu Pro Asp Ser Val Ile Thr Gln Gl - #y Ser Pro Val Thr Leu           50             - #     55             - #     60                          - - Ser Cys Gln Gly Ser Leu Glu Ala Gln Glu Ty - #r Arg Leu Tyr Arg Glu       65                 - # 70                 - # 75                 - # 80       - - Lys Lys Ser Ala Ser Trp Ile Thr Arg Ile Ar - #g Pro Glu Leu Val Lys                       85 - #                 90 - #                 95              - - Asn Gly Gln Phe His Ile Pro Ser Ile Thr Tr - #p Glu His Thr Gly Arg                  100      - #           105      - #           110                  - - Tyr Gly Cys Gln Tyr Tyr Ser Arg Ala Arg Tr - #p Ser Glu Leu Ser Asp              115          - #       120          - #       125                      - - Pro Leu Val Leu Val Met Thr Gly Ala Tyr Pr - #o Lys Pro Thr Leu Ser          130              - #   135              - #   140                          - - Ala Gln Pro Ser Pro Val Val Thr Ser Gly Gl - #y Arg Val Thr Leu Gln      145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - Cys Glu Ser Gln Val Ala Phe Gly Gly Phe Il - #e Leu Cys Lys Glu        Gly                                                                                             165  - #               170  - #               175             - - Glu Asp Glu His Pro Gln Cys Leu Asn Ser Gl - #n Pro His Ala Arg Gly                  180      - #           185      - #           190                  - - Ser Ser Arg Ala Ile Phe Ser Val Gly Pro Va - #l Ser Pro Asn Arg Arg              195          - #       200          - #       205                      - - Trp Ser His Arg Cys Tyr Gly Tyr Asp Leu As - #n Ser Pro Tyr Val Trp          210              - #   215              - #   220                          - - Ser Ser Pro Ser Asp Leu Leu Glu Leu Leu Va - #l Pro Gly Val Ser Lys      225                 2 - #30                 2 - #35                 2 -      #40                                                                              - - Lys Pro Ser Leu Ser Val Gln Pro Gly Pro Va - #l Val Ala Pro Gly        Glu                                                                                             245  - #               250  - #               255             - - Ser Leu Thr Leu Gln Cys Val Ser Asp Val Gl - #y Tyr Asp Arg Phe Val                  260      - #           265      - #           270                  - - Leu Tyr Lys Glu Gly Glu Arg Asp Leu Arg Gl - #n Leu Pro Gly Arg Gln              275          - #       280          - #       285                      - - Pro Gln Ala Gly Leu Ser Gln Ala Asn Phe Th - #r Leu Gly Pro Val Ser          290              - #   295              - #   300                          - - Arg Ser Tyr Gly Gly Gln Tyr Arg Cys Tyr Gl - #y Ala Tyr Asn Leu Ser      305                 3 - #10                 3 - #15                 3 -      #20                                                                              - - Ser Glu Trp Ser Ala Pro Ser Asp Pro Leu As - #p Ile Leu Ile Thr        Gly                                                                                             325  - #               330  - #               335             - - Gln Ile His Gly Thr Pro Phe Ile Ser Val Gl - #n Pro Gly Pro Thr Val                  340      - #           345      - #           350                  - - Ala Ser Gly Glu Asn Val Thr Leu Leu Cys Gl - #n Ser Trp Arg Gln Phe              355          - #       360          - #       365                      - - His Thr Phe Leu Leu Thr Lys Ala Gly Ala Al - #a Asp Ala Pro Leu Arg          370              - #   375              - #   380                          - - Leu Arg Ser Ile His Glu Tyr Pro Lys Tyr Gl - #n Ala Glu Phe Pro Met      385                 3 - #90                 3 - #95                 4 -      #00                                                                              - - Ser Pro Val Thr Ser Ala His Ala Gly Thr Ty - #r Arg Cys Tyr Gly        Ser                                                                                             405  - #               410  - #               415             - - Leu Asn Ser Asp Pro Tyr Leu Leu Ser His Pr - #o Ser Glu Pro Leu Glu                  420      - #           425      - #           430                  - - Leu Val Val Ser Gly Pro Ser Met Gly Ser Se - #r Pro Pro Pro Thr Gly              435          - #       440          - #       445                      - - Pro Ile Ser Thr Pro Ala Gly Pro Glu Asp Gl - #n Pro Leu Thr Pro Thr          450              - #   455              - #   460                          - - Gly Ser Asp Pro Gln Ser Gly Leu Gly Arg Hi - #s Leu Gly Val Val Ile      465                 4 - #70                 4 - #75                 4 -      #80                                                                              - - Gly Ile Leu Val Ala Val Val Leu Leu Leu Le - #u Leu Leu Leu Leu        Leu                                                                                             485  - #               490  - #               495             - - Phe Leu Ile Leu Arg His Arg Arg Gln Gly Ly - #s His Trp Thr Ser Thr                  500      - #           505      - #           510                  - - Gln Arg Lys Ala Asp Phe Gln His Pro Ala Gl - #y Ala Val Gly Pro Glu              515          - #       520          - #       525                      - - Pro Thr Asp Arg Gly Leu Gln Trp Arg Ser Se - #r Pro Ala Ala Asp Ala          530              - #   535              - #   540                          - - Gln Glu Glu Asn Leu Tyr Ala Ala Val Lys As - #p Thr Gln Pro Glu Asp      545                 5 - #50                 5 - #55                 5 -      #60                                                                              - - Gly Val Glu Met Asp Thr Arg Ala Ala Ala Se - #r Glu Ala Pro Gln        Asp                                                                                             565  - #               570  - #               575             - - Val Thr Tyr Ala Gln Leu His Ser Leu Thr Le - #u Arg Arg Lys Ala Thr                  580      - #           585      - #           590                  - - Glu Pro Pro Pro Ser Gln Glu Arg Glu Pro Pr - #o Ala Glu Pro Ser Ile              595          - #       600          - #       605                      - - Tyr Ala Thr Leu Ala Ile His                                                  610              - #   615                                                 - -  - - (2) INFORMATION FOR SEQ ID NO:17:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 2388 base - #pairs                                                (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 180..2024                                              - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:17:                              - - AAAGAAGTCA ACTTTTCTTC CCCTACTTCC CTGCATTTCT CCTCTGTGCT CA -             #CTGCCACA     60                                                                 - - CGCAGCTCAA CCTGGACGGC ACAGCCAGAT GCGAGATGCG TCTCTGCTGA TC -            #TGAGTCTG    120                                                                 - - CCTGCAGCAT GGACCTGGGT CTTCCCTGAA GCATCTCCAG GGCTGGAGGG AC -            #GACTGCC     179                                                                 - - ATG CAC CGA GGG CTC ATC CAT CCG CAG AGC AG - #G GCA GTG GGA GGA        GAC      227                                                                    Met His Arg Gly Leu Ile His Pro Gln Ser Ar - #g Ala Val Gly Gly Asp            1               5 - #                 10 - #                 15              - - GCC ATG ACC CCC ATC GTC ACA GTC CTG ATC TG - #T CTC GGG CTG AGT CTG          275                                                                       Ala Met Thr Pro Ile Val Thr Val Leu Ile Cy - #s Leu Gly Leu Ser Leu                        20     - #             25     - #             30                  - - GGC CCC AGG ACC CAC GTG CAG ACA GGG ACC AT - #C CCC AAG CCC ACC CTG          323                                                                       Gly Pro Arg Thr His Val Gln Thr Gly Thr Il - #e Pro Lys Pro Thr Leu                    35         - #         40         - #         45                      - - TGG GCT GAG CCA GAC TCT GTG ATC ACC CAG GG - #G AGT CCC GTC ACC CTC          371                                                                       Trp Ala Glu Pro Asp Ser Val Ile Thr Gln Gl - #y Ser Pro Val Thr Leu                50             - #     55             - #     60                          - - AGT TGT CAG GGG AGC CTT GAA GCC CAG GAG TA - #C CGT CTA TAT AGG GAG          419                                                                       Ser Cys Gln Gly Ser Leu Glu Ala Gln Glu Ty - #r Arg Leu Tyr Arg Glu            65                 - # 70                 - # 75                 - # 80       - - AAA AAA TCA GCA TCT TGG ATT ACA CGG ATA CG - #A CCA GAG CTT GTG AAG          467                                                                       Lys Lys Ser Ala Ser Trp Ile Thr Arg Ile Ar - #g Pro Glu Leu Val Lys                            85 - #                 90 - #                 95              - - AAC GGC CAG TTC CAC ATC CCA TCC ATC ACC TG - #G GAA CAC ACA GGG CGA          515                                                                       Asn Gly Gln Phe His Ile Pro Ser Ile Thr Tr - #p Glu His Thr Gly Arg                       100      - #           105      - #           110                  - - TAT GGC TGT CAG TAT TAC AGC CGC GCT CGG TG - #G TCT GAG CTC AGT GAC          563                                                                       Tyr Gly Cys Gln Tyr Tyr Ser Arg Ala Arg Tr - #p Ser Glu Leu Ser Asp                   115          - #       120          - #       125                      - - CCC CTG GTG CTG GTG ATG ACA GGA GCC TAC CC - #A AAA CCC ACC CTC TCA          611                                                                       Pro Leu Val Leu Val Met Thr Gly Ala Tyr Pr - #o Lys Pro Thr Leu Ser               130              - #   135              - #   140                          - - GCC CAG CCC AGC CCT GTG GTG ACC TCA GGA GG - #A AGG GTG ACC CTC CAG          659                                                                       Ala Gln Pro Ser Pro Val Val Thr Ser Gly Gl - #y Arg Val Thr Leu Gln           145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - TGT GAG TCA CAG GTG GCA TTT GGC GGC TTC AT - #T CTG TGT AAG GAA        GGA      707                                                                    Cys Glu Ser Gln Val Ala Phe Gly Gly Phe Il - #e Leu Cys Lys Glu Gly                          165  - #               170  - #               175              - - GAA GAT GAA CAC CCA CAA TGC CTG AAC TCC CA - #G CCC CAT GCC CGT GGG          755                                                                       Glu Asp Glu His Pro Gln Cys Leu Asn Ser Gl - #n Pro His Ala Arg Gly                       180      - #           185      - #           190                  - - TCG TCC CGC GCC ATC TTC TCC GTG GGC CCC GT - #G AGC CCG AAT CGC AGG          803                                                                       Ser Ser Arg Ala Ile Phe Ser Val Gly Pro Va - #l Ser Pro Asn Arg Arg                   195          - #       200          - #       205                      - - TGG TCG CAC AGG TGC TAT GGT TAT GAC TTG AA - #C TCT CCC TAT GTG TGG          851                                                                       Trp Ser His Arg Cys Tyr Gly Tyr Asp Leu As - #n Ser Pro Tyr Val Trp               210              - #   215              - #   220                          - - TCT TCA CCC AGT GAT CTC CTG GAG CTC CTG GT - #C CCA GGT GTT TCT AAG          899                                                                       Ser Ser Pro Ser Asp Leu Leu Glu Leu Leu Va - #l Pro Gly Val Ser Lys           225                 2 - #30                 2 - #35                 2 -      #40                                                                              - - AAG CCA TCA CTC TCA GTG CAG CCG GGT CCT GT - #C GTG GCC CCT GGG        GAA      947                                                                    Lys Pro Ser Leu Ser Val Gln Pro Gly Pro Va - #l Val Ala Pro Gly Glu                          245  - #               250  - #               255              - - AGC CTG ACC CTC CAG TGT GTC TCT GAT GTC GG - #C TAT GAC AGA TTT GTT          995                                                                       Ser Leu Thr Leu Gln Cys Val Ser Asp Val Gl - #y Tyr Asp Arg Phe Val                       260      - #           265      - #           270                  - - CTG TAC AAG GAG GGG GAA CGT GAC CTT CGC CA - #G CTC CCT GGC CGG CAG         1043                                                                       Leu Tyr Lys Glu Gly Glu Arg Asp Leu Arg Gl - #n Leu Pro Gly Arg Gln                   275          - #       280          - #       285                      - - CCC CAG GCT GGG CTC TCC CAG GCC AAC TTC AC - #C CTG GGC CCT GTG AGC         1091                                                                       Pro Gln Ala Gly Leu Ser Gln Ala Asn Phe Th - #r Leu Gly Pro Val Ser               290              - #   295              - #   300                          - - CGC TCC TAC GGG GGC CAG TAC AGA TGC TAC GG - #T GCA TAC AAC CTC TCC         1139                                                                       Arg Ser Tyr Gly Gly Gln Tyr Arg Cys Tyr Gl - #y Ala Tyr Asn Leu Ser           305                 3 - #10                 3 - #15                 3 -      #20                                                                              - - TCC GAG TGG TCG GCC CCC AGC GAC CCC CTG GA - #C ATC CTG ATC ACA        GGA     1187                                                                    Ser Glu Trp Ser Ala Pro Ser Asp Pro Leu As - #p Ile Leu Ile Thr Gly                          325  - #               330  - #               335              - - CAG ATC CAT GGC ACA CCC TTC ATC TCA GTG CA - #G CCA GGC CCC ACA GTG         1235                                                                       Gln Ile His Gly Thr Pro Phe Ile Ser Val Gl - #n Pro Gly Pro Thr Val                       340      - #           345      - #           350                  - - GCC TCA GGA GAG AAC GTG ACC CTG CTG TGT CA - #G TCA TGG CGG CAG TTC         1283                                                                       Ala Ser Gly Glu Asn Val Thr Leu Leu Cys Gl - #n Ser Trp Arg Gln Phe                   355          - #       360          - #       365                      - - CAC ACT TTC CTT CTG ACC AAG GCG GGA GCA GC - #T GAT GCC CCA CTC CGT         1331                                                                       His Thr Phe Leu Leu Thr Lys Ala Gly Ala Al - #a Asp Ala Pro Leu Arg               370              - #   375              - #   380                          - - CTA AGA TCA ATA CAC GAA TAT CCT AAG TAC CA - #G GCT GAA TTC CCC ATG         1379                                                                       Leu Arg Ser Ile His Glu Tyr Pro Lys Tyr Gl - #n Ala Glu Phe Pro Met           385                 3 - #90                 3 - #95                 4 -      #00                                                                              - - AGT CCC GTG ACC TCA GCC CAC GCG GGG ACC TA - #C AGG TGC TAC GGC        TCA     1427                                                                    Ser Pro Val Thr Ser Ala His Ala Gly Thr Ty - #r Arg Cys Tyr Gly Ser                          405  - #               410  - #               415              - - CTC AAC TCC GAC CCC TAC CTG CTG TCT CAC CC - #C AGT GAG CCC CTG GAG         1475                                                                       Leu Asn Ser Asp Pro Tyr Leu Leu Ser His Pr - #o Ser Glu Pro Leu Glu                       420      - #           425      - #           430                  - - CTC GTG GTC TCA GGA CCC TCC ATG GGT TCC AG - #C CCC CCA CCC ACC GGT         1523                                                                       Leu Val Val Ser Gly Pro Ser Met Gly Ser Se - #r Pro Pro Pro Thr Gly                   435          - #       440          - #       445                      - - CCC ATC TCC ACA CCT GCA GGC CCT GAG GAC CA - #G CCC CTC ACC CCC ACT         1571                                                                       Pro Ile Ser Thr Pro Ala Gly Pro Glu Asp Gl - #n Pro Leu Thr Pro Thr               450              - #   455              - #   460                          - - GGG TCG GAT CCC CAA AGT GGT CTG GGA AGG CA - #C CTG GGG GTT GTG ATC         1619                                                                       Gly Ser Asp Pro Gln Ser Gly Leu Gly Arg Hi - #s Leu Gly Val Val Ile           465                 4 - #70                 4 - #75                 4 -      #80                                                                              - - GGC ATC TTG GTG GCC GTC GTC CTA CTG CTC CT - #C CTC CTC CTC CTC        CTC     1667                                                                    Gly Ile Leu Val Ala Val Val Leu Leu Leu Le - #u Leu Leu Leu Leu Leu                          485  - #               490  - #               495              - - TTC CTC ATC CTC CGA CAT CGA CGT CAG GGC AA - #A CAC TGG ACA TCG ACC         1715                                                                       Phe Leu Ile Leu Arg His Arg Arg Gln Gly Ly - #s His Trp Thr Ser Thr                       500      - #           505      - #           510                  - - CAG AGA AAG GCT GAT TTC CAA CAT CCT GCA GG - #G GCT GTG GGG CCA GAG         1763                                                                       Gln Arg Lys Ala Asp Phe Gln His Pro Ala Gl - #y Ala Val Gly Pro Glu                   515          - #       520          - #       525                      - - CCC ACA GAC AGA GGC CTG CAG TGG AGG TCC AG - #C CCA GCT GCC GAC GCC         1811                                                                       Pro Thr Asp Arg Gly Leu Gln Trp Arg Ser Se - #r Pro Ala Ala Asp Ala               530              - #   535              - #   540                          - - CAG GAA GAA AAC CTC TAT GCT GCC GTG AAG GA - #C ACA CAG CCT GAA GAT         1859                                                                       Gln Glu Glu Asn Leu Tyr Ala Ala Val Lys As - #p Thr Gln Pro Glu Asp           545                 5 - #50                 5 - #55                 5 -      #60                                                                              - - GGG GTG GAG ATG GAC ACT CGG GCT GCT GCA TC - #T GAA GCC CCC CAG        GAT     1907                                                                    Gly Val Glu Met Asp Thr Arg Ala Ala Ala Se - #r Glu Ala Pro Gln Asp                          565  - #               570  - #               575              - - GTG ACC TAC GCC CAG CTG CAC AGC TTG ACC CT - #C AGA CGG AAG GCA ACT         1955                                                                       Val Thr Tyr Ala Gln Leu His Ser Leu Thr Le - #u Arg Arg Lys Ala Thr                       580      - #           585      - #           590                  - - GAG CCT CCT CCA TCC CAG GAA AGG GAA CCT CC - #A GCT GAG CCC AGC ATC         2003                                                                       Glu Pro Pro Pro Ser Gln Glu Arg Glu Pro Pr - #o Ala Glu Pro Ser Ile                   595          - #       600          - #       605                      - - TAC GCC ACC CTG GCC ATC CAC TAGCCCGGAG GGTACGCAG - #A CTCCACACTC            2054                                                                       Tyr Ala Thr Leu Ala Ile His                                                       610              - #   615                                                 - - AGTAGAAGGA GACTCAGGAC TGCTGAAGGC ACGGGAGCTG CCCCCAGTGG AC -             #ACCAATGA   2114                                                                 - - ACCCCAGTCA GCCTGGACCC CTAACAAAGA CCATGAGGAG ATGCTGGGAA CT -            #TTGGGACT   2174                                                                 - - CACTTGATTC TGCAGTCGAA ATAACTAATA TCCCTACATT TTTTAATTAA AG -            #CAACAGAC   2234                                                                 - - TTCTCAATAA TCAATGAGTT AACCGAGAAA ACTAAAATCA GAAGTAAGAA TG -            #TGCTTTAA   2294                                                                 - - ACTGAATCAC AATATAAATA TTACACATCA CACAATGAAA TTGAAAAAGT AC -            #AAACCACA   2354                                                                 - - AATGAAAAAA GTAGAAACGA AAAAAAAAAA AAAA       - #                  -     #      2388                                                                     - -  - - (2) INFORMATION FOR SEQ ID NO:18:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 615 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:18:                              - - Met His Arg Gly Leu Ile His Pro Gln Ser Ar - #g Ala Val Gly Gly Asp        1               5 - #                 10 - #                 15              - - Ala Met Thr Pro Ile Val Thr Val Leu Ile Cy - #s Leu Gly Leu Ser Leu                   20     - #             25     - #             30                  - - Gly Pro Arg Thr His Val Gln Thr Gly Thr Il - #e Pro Lys Pro Thr Leu               35         - #         40         - #         45                      - - Trp Ala Glu Pro Asp Ser Val Ile Thr Gln Gl - #y Ser Pro Val Thr Leu           50             - #     55             - #     60                          - - Ser Cys Gln Gly Ser Leu Glu Ala Gln Glu Ty - #r Arg Leu Tyr Arg Glu       65                 - # 70                 - # 75                 - # 80       - - Lys Lys Ser Ala Ser Trp Ile Thr Arg Ile Ar - #g Pro Glu Leu Val Lys                       85 - #                 90 - #                 95              - - Asn Gly Gln Phe His Ile Pro Ser Ile Thr Tr - #p Glu His Thr Gly Arg                  100      - #           105      - #           110                  - - Tyr Gly Cys Gln Tyr Tyr Ser Arg Ala Arg Tr - #p Ser Glu Leu Ser Asp              115          - #       120          - #       125                      - - Pro Leu Val Leu Val Met Thr Gly Ala Tyr Pr - #o Lys Pro Thr Leu Ser          130              - #   135              - #   140                          - - Ala Gln Pro Ser Pro Val Val Thr Ser Gly Gl - #y Arg Val Thr Leu Gln      145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - Cys Glu Ser Gln Val Ala Phe Gly Gly Phe Il - #e Leu Cys Lys Glu        Gly                                                                                             165  - #               170  - #               175             - - Glu Asp Glu His Pro Gln Cys Leu Asn Ser Gl - #n Pro His Ala Arg Gly                  180      - #           185      - #           190                  - - Ser Ser Arg Ala Ile Phe Ser Val Gly Pro Va - #l Ser Pro Asn Arg Arg              195          - #       200          - #       205                      - - Trp Ser His Arg Cys Tyr Gly Tyr Asp Leu As - #n Ser Pro Tyr Val Trp          210              - #   215              - #   220                          - - Ser Ser Pro Ser Asp Leu Leu Glu Leu Leu Va - #l Pro Gly Val Ser Lys      225                 2 - #30                 2 - #35                 2 -      #40                                                                              - - Lys Pro Ser Leu Ser Val Gln Pro Gly Pro Va - #l Val Ala Pro Gly        Glu                                                                                             245  - #               250  - #               255             - - Ser Leu Thr Leu Gln Cys Val Ser Asp Val Gl - #y Tyr Asp Arg Phe Val                  260      - #           265      - #           270                  - - Leu Tyr Lys Glu Gly Glu Arg Asp Leu Arg Gl - #n Leu Pro Gly Arg Gln              275          - #       280          - #       285                      - - Pro Gln Ala Gly Leu Ser Gln Ala Asn Phe Th - #r Leu Gly Pro Val Ser          290              - #   295              - #   300                          - - Arg Ser Tyr Gly Gly Gln Tyr Arg Cys Tyr Gl - #y Ala Tyr Asn Leu Ser      305                 3 - #10                 3 - #15                 3 -      #20                                                                              - - Ser Glu Trp Ser Ala Pro Ser Asp Pro Leu As - #p Ile Leu Ile Thr        Gly                                                                                             325  - #               330  - #               335             - - Gln Ile His Gly Thr Pro Phe Ile Ser Val Gl - #n Pro Gly Pro Thr Val                  340      - #           345      - #           350                  - - Ala Ser Gly Glu Asn Val Thr Leu Leu Cys Gl - #n Ser Trp Arg Gln Phe              355          - #       360          - #       365                      - - His Thr Phe Leu Leu Thr Lys Ala Gly Ala Al - #a Asp Ala Pro Leu Arg          370              - #   375              - #   380                          - - Leu Arg Ser Ile His Glu Tyr Pro Lys Tyr Gl - #n Ala Glu Phe Pro Met      385                 3 - #90                 3 - #95                 4 -      #00                                                                              - - Ser Pro Val Thr Ser Ala His Ala Gly Thr Ty - #r Arg Cys Tyr Gly        Ser                                                                                             405  - #               410  - #               415             - - Leu Asn Ser Asp Pro Tyr Leu Leu Ser His Pr - #o Ser Glu Pro Leu Glu                  420      - #           425      - #           430                  - - Leu Val Val Ser Gly Pro Ser Met Gly Ser Se - #r Pro Pro Pro Thr Gly              435          - #       440          - #       445                      - - Pro Ile Ser Thr Pro Ala Gly Pro Glu Asp Gl - #n Pro Leu Thr Pro Thr          450              - #   455              - #   460                          - - Gly Ser Asp Pro Gln Ser Gly Leu Gly Arg Hi - #s Leu Gly Val Val Ile      465                 4 - #70                 4 - #75                 4 -      #80                                                                              - - Gly Ile Leu Val Ala Val Val Leu Leu Leu Le - #u Leu Leu Leu Leu        Leu                                                                                             485  - #               490  - #               495             - - Phe Leu Ile Leu Arg His Arg Arg Gln Gly Ly - #s His Trp Thr Ser Thr                  500      - #           505      - #           510                  - - Gln Arg Lys Ala Asp Phe Gln His Pro Ala Gl - #y Ala Val Gly Pro Glu              515          - #       520          - #       525                      - - Pro Thr Asp Arg Gly Leu Gln Trp Arg Ser Se - #r Pro Ala Ala Asp Ala          530              - #   535              - #   540                          - - Gln Glu Glu Asn Leu Tyr Ala Ala Val Lys As - #p Thr Gln Pro Glu Asp      545                 5 - #50                 5 - #55                 5 -      #60                                                                              - - Gly Val Glu Met Asp Thr Arg Ala Ala Ala Se - #r Glu Ala Pro Gln        Asp                                                                                             565  - #               570  - #               575             - - Val Thr Tyr Ala Gln Leu His Ser Leu Thr Le - #u Arg Arg Lys Ala Thr                  580      - #           585      - #           590                  - - Glu Pro Pro Pro Ser Gln Glu Arg Glu Pro Pr - #o Ala Glu Pro Ser Ile              595          - #       600          - #       605                      - - Tyr Ala Thr Leu Ala Ile His                                                  610              - #   615                                                 - -  - - (2) INFORMATION FOR SEQ ID NO:19:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 2200 base - #pairs                                                (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 174..1466                                              - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:19:                              - - GTCAACTTTT CTTCCCCTAC TTCCCTGCAT TTCTCCTCTG TGCTCACTGC CA -             #CACGCAGC     60                                                                 - - TCAACCTGGA CGGCACAGCC AGATGCGAGA TGCGTCTCTG CTGATCTGAG TC -            #TGCCTGCA    120                                                                 - - GCATGGACCT GGGTCTTCCC TGAAGCATCT CCAGGGCTGG AGGGACGACT GC - #C ATG           176                                                                                        - #                  - #                  - #     Met                         - #                  - #                  - #       1        - - CAC CGA GGG CTC ATC CAT CCG CAG AGC AGG GC - #A GTG GGA GGA GAC GCC          224                                                                       His Arg Gly Leu Ile His Pro Gln Ser Arg Al - #a Val Gly Gly Asp Ala                         5    - #              10    - #              15                  - - ATG ACC CCC ATC GTC ACA GTC CTG ATC TGT CT - #C GGG CTG AGT CTG GGC          272                                                                       Met Thr Pro Ile Val Thr Val Leu Ile Cys Le - #u Gly Leu Ser Leu Gly                    20         - #         25         - #         30                      - - CCC AGG ACC CAC GTG CAG ACA GGG ACC ATC CC - #C AAG CCC ACC CTG TGG          320                                                                       Pro Arg Thr His Val Gln Thr Gly Thr Ile Pr - #o Lys Pro Thr Leu Trp                35             - #     40             - #     45                          - - GCT GAG CCA GAC TCT GTG ATC ACC CAG GGG AG - #T CCC GTC ACC CTC AGT          368                                                                       Ala Glu Pro Asp Ser Val Ile Thr Gln Gly Se - #r Pro Val Thr Leu Ser            50                 - # 55                 - # 60                 - # 65       - - TGT CAG GGG AGC CTT GAA GCC CAG GAG TAC CG - #T CTA TAT AGG GAG AAA          416                                                                       Cys Gln Gly Ser Leu Glu Ala Gln Glu Tyr Ar - #g Leu Tyr Arg Glu Lys                            70 - #                 75 - #                 80              - - AAA TCA GCA TCT TGG ATT ACA CGG ATA CGA CC - #A GAG CTT GTG AAG AAC          464                                                                       Lys Ser Ala Ser Trp Ile Thr Arg Ile Arg Pr - #o Glu Leu Val Lys Asn                        85     - #             90     - #             95                  - - GGC CAG TTC CAC ATC CCA TCC ATC ACC TGG GA - #A CAC ACA GGG CGA TAT          512                                                                       Gly Gln Phe His Ile Pro Ser Ile Thr Trp Gl - #u His Thr Gly Arg Tyr                   100          - #       105          - #       110                      - - GGC TGT CAG TAT TAC AGC CGC GCT CGG TGG TC - #T GAG CTC AGT GAC CCC          560                                                                       Gly Cys Gln Tyr Tyr Ser Arg Ala Arg Trp Se - #r Glu Leu Ser Asp Pro               115              - #   120              - #   125                          - - CTG GTG CTG GTG ATG ACA GGA GCC TAC CCA AA - #A CCC ACC CTC TCA GCC          608                                                                       Leu Val Leu Val Met Thr Gly Ala Tyr Pro Ly - #s Pro Thr Leu Ser Ala           130                 1 - #35                 1 - #40                 1 -      #45                                                                              - - CAG CCC AGC CCT GTG GTG ACC TCA GGA GGA AG - #G GTG ACC CTC CAG        TGT      656                                                                    Gln Pro Ser Pro Val Val Thr Ser Gly Gly Ar - #g Val Thr Leu Gln Cys                          150  - #               155  - #               160              - - GAG TCA CAG GTG GCA TTT GGC GGC TTC ATT CT - #G TGT AAG GAA GGA GAA          704                                                                       Glu Ser Gln Val Ala Phe Gly Gly Phe Ile Le - #u Cys Lys Glu Gly Glu                       165      - #           170      - #           175                  - - GAT GAA CAC CCA CAA TGC CTG AAC TCC CAG CC - #C CAT GCC CGT GGG TCG          752                                                                       Asp Glu His Pro Gln Cys Leu Asn Ser Gln Pr - #o His Ala Arg Gly Ser                   180          - #       185          - #       190                      - - TCC CGC GCC ATC TTC TCC GTG GGC CCC GTG AG - #C CCG AAT CGC AGG TGG          800                                                                       Ser Arg Ala Ile Phe Ser Val Gly Pro Val Se - #r Pro Asn Arg Arg Trp               195              - #   200              - #   205                          - - TCG CAC AGG TGC TAT GGT TAT GAC TTG AAC TC - #T CCC TAT GTG TGG TCT          848                                                                       Ser His Arg Cys Tyr Gly Tyr Asp Leu Asn Se - #r Pro Tyr Val Trp Ser           210                 2 - #15                 2 - #20                 2 -      #25                                                                              - - TCA CCC AGT GAT CTC CTG GAG CTC CTG GTC CC - #A GGT GTT TCT AAG        AAG      896                                                                    Ser Pro Ser Asp Leu Leu Glu Leu Leu Val Pr - #o Gly Val Ser Lys Lys                          230  - #               235  - #               240              - - CCA TCA CTC TCA GTG CAG CCG GGT CCT GTC GT - #G GCC CCT GGG GAA AGC          944                                                                       Pro Ser Leu Ser Val Gln Pro Gly Pro Val Va - #l Ala Pro Gly Glu Ser                       245      - #           250      - #           255                  - - CTG ACC CTC CAG TGT GTC TCT GAT GTC GGC TA - #T GAC AGA TTT GTT CTG          992                                                                       Leu Thr Leu Gln Cys Val Ser Asp Val Gly Ty - #r Asp Arg Phe Val Leu                   260          - #       265          - #       270                      - - TAC AAG GAG GGG GAA CGT GAC CTT CGC CAG CT - #C CCT GGC CGG CAG CCC         1040                                                                       Tyr Lys Glu Gly Glu Arg Asp Leu Arg Gln Le - #u Pro Gly Arg Gln Pro               275              - #   280              - #   285                          - - CAG GCT GGG CTC TCC CAG GCC AAC TTC ACC CT - #G GGC CCT GTG AGC CGC         1088                                                                       Gln Ala Gly Leu Ser Gln Ala Asn Phe Thr Le - #u Gly Pro Val Ser Arg           290                 2 - #95                 3 - #00                 3 -      #05                                                                              - - TCC TAC GGG GGC CAG TAC AGA TGC TAC GGT GC - #A TAC AAC CTC TCC        TCC     1136                                                                    Ser Tyr Gly Gly Gln Tyr Arg Cys Tyr Gly Al - #a Tyr Asn Leu Ser Ser                          310  - #               315  - #               320              - - GAG TGG TCG GCC CCC AGC GAC CCC CTG GAC AT - #C CTG ATC ACA GGA CAG         1184                                                                       Glu Trp Ser Ala Pro Ser Asp Pro Leu Asp Il - #e Leu Ile Thr Gly Gln                       325      - #           330      - #           335                  - - ATC CAT GGC ACA CCC TTC ATC TCA GTG CAG CC - #A GGC CCC ACA GTG GCC         1232                                                                       Ile His Gly Thr Pro Phe Ile Ser Val Gln Pr - #o Gly Pro Thr Val Ala                   340          - #       345          - #       350                      - - TCA GGA GAG AAC GTG ACC CTG CTG TGT CAG TC - #A TGG CGG CAG TTC CAC         1280                                                                       Ser Gly Glu Asn Val Thr Leu Leu Cys Gln Se - #r Trp Arg Gln Phe His               355              - #   360              - #   365                          - - ACT TTC CTT CTG ACC AAG GCG GGA GCA GCT GA - #T GCC CCA CTC CGT CTA         1328                                                                       Thr Phe Leu Leu Thr Lys Ala Gly Ala Ala As - #p Ala Pro Leu Arg Leu           370                 3 - #75                 3 - #80                 3 -      #85                                                                              - - AGA TCA ATA CAC GAA TAT CCT AAG TAC CAG GC - #T GAA TTC CCC ATG        AGT     1376                                                                    Arg Ser Ile His Glu Tyr Pro Lys Tyr Gln Al - #a Glu Phe Pro Met Ser                          390  - #               395  - #               400              - - CCT GTG ACC TCA GCC CAC GCG GGG ACC TAC AG - #G ACC CTC CAT GGG TTC         1424                                                                       Pro Val Thr Ser Ala His Ala Gly Thr Tyr Ar - #g Thr Leu His Gly Phe                       405      - #           410      - #           415                  - - CAG CCC CCC ACC CAC CGG TCC CAT CTC CAC AC - #C TGC AGG CCC                 - #1466                                                                    Gln Pro Pro Thr His Arg Ser His Leu His Th - #r Cys Arg Pro                           420          - #       425          - #       430                      - - TGAGGACCAG CCCCTCACCC CCACTGGGTC GGATCCCCAA AGTGGTCTGG GA -             #AGGCACCT   1526                                                                 - - GGGGGTTGTG ATCGGCATCT TGGTGGCCGT CGTCCTACTG CTCCTCCTCC TC -            #CTCCTCCT   1586                                                                 - - CTTCCTCATC CTCCGACATC GACGTCAGGG CAAACACTGG ACATCGACCC AG -            #AGAAAGGC   1646                                                                 - - TGATTTCCAA CATCCTGCAG GGGCTGTGGG GCCAGAGCCC ACAGACAGAG GC -            #CTGCAGTG   1706                                                                 - - GAGGTCCAGC CCAGCTGCCG ACGCCCAGGA AGAAAACCTC TATGCTGCCG TG -            #AAGGACAC   1766                                                                 - - ACAGCCTGAA GATGGGGTGG AGATGGACAC TCGGGCTGCT GCATCTGAAG CC -            #CCCCAGGA   1826                                                                 - - TGTGACCTAC GCCCAGCTGC ACAGCTTGAC CCTCAGACGG AAGGCAACTG AG -            #CCTCCTCC   1886                                                                 - - ATCCCAGGAA AGGGAACCTC CAGCTGAGCC CAGCATCTAC GCCACCCTGG CC -            #ATCCACTA   1946                                                                 - - GCCCGGAGGG TACGCAGACT CCACACTCAG TAGAAGGAGA CTCAGGACTG CT -            #GAAGGCAC   2006                                                                 - - GGGAGCTGCC CCCAGTGGAC ACCAATGAAC CCCAGTCAGC CTGGACCCCT AA -            #CAAAGACC   2066                                                                 - - ATGAGGAGAT GCTGGGAACT TTGGGACTCA CTTGATTCTG CAGTCGAAAT AA -            #CTAATATC   2126                                                                 - - CCTACATTTT TTAATTAAAG CAACAGACTT CTCAATAATC AATGAGTTAA CC -            #GAGAAAAC   2186                                                                 - - TAAAAAAAAA AAAA              - #                  - #                      - #   2200                                                                  - -  - - (2) INFORMATION FOR SEQ ID NO:20:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 431 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:20:                              - - Met His Arg Gly Leu Ile His Pro Gln Ser Ar - #g Ala Val Gly Gly Asp        1               5 - #                 10 - #                 15              - - Ala Met Thr Pro Ile Val Thr Val Leu Ile Cy - #s Leu Gly Leu Ser Leu                   20     - #             25     - #             30                  - - Gly Pro Arg Thr His Val Gln Thr Gly Thr Il - #e Pro Lys Pro Thr Leu               35         - #         40         - #         45                      - - Trp Ala Glu Pro Asp Ser Val Ile Thr Gln Gl - #y Ser Pro Val Thr Leu           50             - #     55             - #     60                          - - Ser Cys Gln Gly Ser Leu Glu Ala Gln Glu Ty - #r Arg Leu Tyr Arg Glu       65                 - # 70                 - # 75                 - # 80       - - Lys Lys Ser Ala Ser Trp Ile Thr Arg Ile Ar - #g Pro Glu Leu Val Lys                       85 - #                 90 - #                 95              - - Asn Gly Gln Phe His Ile Pro Ser Ile Thr Tr - #p Glu His Thr Gly Arg                  100      - #           105      - #           110                  - - Tyr Gly Cys Gln Tyr Tyr Ser Arg Ala Arg Tr - #p Ser Glu Leu Ser Asp              115          - #       120          - #       125                      - - Pro Leu Val Leu Val Met Thr Gly Ala Tyr Pr - #o Lys Pro Thr Leu Ser          130              - #   135              - #   140                          - - Ala Gln Pro Ser Pro Val Val Thr Ser Gly Gl - #y Arg Val Thr Leu Gln      145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - Cys Glu Ser Gln Val Ala Phe Gly Gly Phe Il - #e Leu Cys Lys Glu        Gly                                                                                             165  - #               170  - #               175             - - Glu Asp Glu His Pro Gln Cys Leu Asn Ser Gl - #n Pro His Ala Arg Gly                  180      - #           185      - #           190                  - - Ser Ser Arg Ala Ile Phe Ser Val Gly Pro Va - #l Ser Pro Asn Arg Arg              195          - #       200          - #       205                      - - Trp Ser His Arg Cys Tyr Gly Tyr Asp Leu As - #n Ser Pro Tyr Val Trp          210              - #   215              - #   220                          - - Ser Ser Pro Ser Asp Leu Leu Glu Leu Leu Va - #l Pro Gly Val Ser Lys      225                 2 - #30                 2 - #35                 2 -      #40                                                                              - - Lys Pro Ser Leu Ser Val Gln Pro Gly Pro Va - #l Val Ala Pro Gly        Glu                                                                                             245  - #               250  - #               255             - - Ser Leu Thr Leu Gln Cys Val Ser Asp Val Gl - #y Tyr Asp Arg Phe Val                  260      - #           265      - #           270                  - - Leu Tyr Lys Glu Gly Glu Arg Asp Leu Arg Gl - #n Leu Pro Gly Arg Gln              275          - #       280          - #       285                      - - Pro Gln Ala Gly Leu Ser Gln Ala Asn Phe Th - #r Leu Gly Pro Val Ser          290              - #   295              - #   300                          - - Arg Ser Tyr Gly Gly Gln Tyr Arg Cys Tyr Gl - #y Ala Tyr Asn Leu Ser      305                 3 - #10                 3 - #15                 3 -      #20                                                                              - - Ser Glu Trp Ser Ala Pro Ser Asp Pro Leu As - #p Ile Leu Ile Thr        Gly                                                                                             325  - #               330  - #               335             - - Gln Ile His Gly Thr Pro Phe Ile Ser Val Gl - #n Pro Gly Pro Thr Val                  340      - #           345      - #           350                  - - Ala Ser Gly Glu Asn Val Thr Leu Leu Cys Gl - #n Ser Trp Arg Gln Phe              355          - #       360          - #       365                      - - His Thr Phe Leu Leu Thr Lys Ala Gly Ala Al - #a Asp Ala Pro Leu Arg          370              - #   375              - #   380                          - - Leu Arg Ser Ile His Glu Tyr Pro Lys Tyr Gl - #n Ala Glu Phe Pro Met      385                 3 - #90                 3 - #95                 4 -      #00                                                                              - - Ser Pro Val Thr Ser Ala His Ala Gly Thr Ty - #r Arg Thr Leu His        Gly                                                                                             405  - #               410  - #               415             - - Phe Gln Pro Pro Thr His Arg Ser His Leu Hi - #s Thr Cys Arg Pro                      420      - #           425      - #           430                  - -  - - (2) INFORMATION FOR SEQ ID NO:21:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 2790 base - #pairs                                                (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 177..2132                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: misc.sub.-- - #feature                                          (B) LOCATION: 1722                                                            (D) OTHER INFORMATION: - #/note= "nucleotide 1722 designated                       C, may - #be A, C, G, or T"                                     - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:21:                              - - GCCACACGCA GCTCAGCCTG GGCGGCACAG CCAGATGCGA GATGCGTCTC TG -             #CTGATCTG     60                                                                 - - AGTCTGCCTG CAGCATGGAC CTGGGTCTTC CCTGAAGCAT CTCCAGGGCT GG -            #AGGGACGA    120                                                                 - - CTGCCATGCA CCGAGGGCTC ATCCATCCAC AGAGCAGGGC AGTGGGAGGA GA - #CGCC            176                                                                       - - ATG ACC CCC ATC CTC ACG GTC CTG ATC TGT CT - #C GGG CTG AGT CTG GGC          224                                                                       Met Thr Pro Ile Leu Thr Val Leu Ile Cys Le - #u Gly Leu Ser Leu Gly             1               5 - #                 10 - #                 15              - - CCC CGG ACC CAC GTG CAG GCA GGG CAC CTC CC - #C AAG CCC ACC CTC TGG          272                                                                       Pro Arg Thr His Val Gln Ala Gly His Leu Pr - #o Lys Pro Thr Leu Trp                        20     - #             25     - #             30                  - - GCT GAA CCA GGC TCT GTG ATC ACC CAG GGG AG - #T CCT GTG ACC CTC AGG          320                                                                       Ala Glu Pro Gly Ser Val Ile Thr Gln Gly Se - #r Pro Val Thr Leu Arg                    35         - #         40         - #         45                      - - TGT CAG GGG GGC CAG GAG ACC CAG GAG TAC CG - #T CTA TAT AGA GAA AAG          368                                                                       Cys Gln Gly Gly Gln Glu Thr Gln Glu Tyr Ar - #g Leu Tyr Arg Glu Lys                50             - #     55             - #     60                          - - AAA ACA GCA CCC TGG ATT ACA CGG ATC CCA CA - #G GAG CTT GTG AAG AAG          416                                                                       Lys Thr Ala Pro Trp Ile Thr Arg Ile Pro Gl - #n Glu Leu Val Lys Lys            65                 - # 70                 - # 75                 - # 80       - - GGC CAG TTC CCC ATC CCA TCC ATC ACC TGG GA - #A CAT GCA GGG CGG TAT          464                                                                       Gly Gln Phe Pro Ile Pro Ser Ile Thr Trp Gl - #u His Ala Gly Arg Tyr                            85 - #                 90 - #                 95              - - CGC TGT TAC TAT GGT AGC GAC ACT GCA GGC CG - #C TCA GAG AGC AGT GAC          512                                                                       Arg Cys Tyr Tyr Gly Ser Asp Thr Ala Gly Ar - #g Ser Glu Ser Ser Asp                       100      - #           105      - #           110                  - - CCC CTG GAG CTG GTG GTG ACA GGA GCC TAC AT - #C AAA CCC ACC CTC TCA          560                                                                       Pro Leu Glu Leu Val Val Thr Gly Ala Tyr Il - #e Lys Pro Thr Leu Ser                   115          - #       120          - #       125                      - - GCC CAG CCC AGC CCC GTG GTG AAC TCA GGA GG - #G AAT GTA ACC CTC CAG          608                                                                       Ala Gln Pro Ser Pro Val Val Asn Ser Gly Gl - #y Asn Val Thr Leu Gln               130              - #   135              - #   140                          - - TGT GAC TCA CAG GTG GCA TTT GAT GGC TTC AT - #T CTG TGT AAG GAA GGA          656                                                                       Cys Asp Ser Gln Val Ala Phe Asp Gly Phe Il - #e Leu Cys Lys Glu Gly           145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - GAA GAT GAA CAC CCA CAA TGC CTG AAC TCC CA - #G CCC CAT GCC CGT        GGG      704                                                                    Glu Asp Glu His Pro Gln Cys Leu Asn Ser Gl - #n Pro His Ala Arg Gly                          165  - #               170  - #               175              - - TCG TCC CGC GCC ATC TTC TCC GTG GGC CCC GT - #G AGC CCG AGT CGC AGG          752                                                                       Ser Ser Arg Ala Ile Phe Ser Val Gly Pro Va - #l Ser Pro Ser Arg Arg                       180      - #           185      - #           190                  - - TGG TGG TAC AGG TGC TAT GCT TAT GAC TCG AA - #C TCT CCC TAT GAG TGG          800                                                                       Trp Trp Tyr Arg Cys Tyr Ala Tyr Asp Ser As - #n Ser Pro Tyr Glu Trp                   195          - #       200          - #       205                      - - TCT CTA CCC AGT GAT CTC CTG GAG CTC CTG GT - #C CTA GGT GTT TCT AAG          848                                                                       Ser Leu Pro Ser Asp Leu Leu Glu Leu Leu Va - #l Leu Gly Val Ser Lys               210              - #   215              - #   220                          - - AAG CCA TCA CTC TCA GTG CAG CCA GGT CCT AT - #C GTG GCC CCT GAG GAG          896                                                                       Lys Pro Ser Leu Ser Val Gln Pro Gly Pro Il - #e Val Ala Pro Glu Glu           225                 2 - #30                 2 - #35                 2 -      #40                                                                              - - ACC CTG ACT CTG CAG TGT GGC TCT GAT GCT GG - #C TAC AAC AGA TTT        GTT      944                                                                    Thr Leu Thr Leu Gln Cys Gly Ser Asp Ala Gl - #y Tyr Asn Arg Phe Val                          245  - #               250  - #               255              - - CTG TAT AAG GAC GGG GAA CGT GAC TTC CTT CA - #G CTC GCT GGC GCA CAG          992                                                                       Leu Tyr Lys Asp Gly Glu Arg Asp Phe Leu Gl - #n Leu Ala Gly Ala Gln                       260      - #           265      - #           270                  - - CCC CAG GCT GGG CTC TCC CAG GCC AAC TTC AC - #C CTG GGC CCT GTG AGC         1040                                                                       Pro Gln Ala Gly Leu Ser Gln Ala Asn Phe Th - #r Leu Gly Pro Val Ser                   275          - #       280          - #       285                      - - CGC TCC TAC GGG GGC CAG TAC AGA TGC TAC GG - #T GCA CAC AAC CTC TCC         1088                                                                       Arg Ser Tyr Gly Gly Gln Tyr Arg Cys Tyr Gl - #y Ala His Asn Leu Ser               290              - #   295              - #   300                          - - TCC GAG TGG TCG GCC CCC AGC GAC CCC CTG GA - #C ATC CTG ATC GCA GGA         1136                                                                       Ser Glu Trp Ser Ala Pro Ser Asp Pro Leu As - #p Ile Leu Ile Ala Gly           305                 3 - #10                 3 - #15                 3 -      #20                                                                              - - CAG TTC TAT GAC AGA GTC TCC CTC TCG GTG CA - #G CCG GGC CCC ACG        GTG     1184                                                                    Gln Phe Tyr Asp Arg Val Ser Leu Ser Val Gl - #n Pro Gly Pro Thr Val                          325  - #               330  - #               335              - - GCC TCA GGA GAG AAC GTG ACC CTG CTG TGT CA - #G TCA CAG GGA TGG ATG         1232                                                                       Ala Ser Gly Glu Asn Val Thr Leu Leu Cys Gl - #n Ser Gln Gly Trp Met                       340      - #           345      - #           350                  - - CAA ACT TTC CTT CTG ACC AAG GAG GGG GCA GC - #T GAT GAC CCA TGG CGT         1280                                                                       Gln Thr Phe Leu Leu Thr Lys Glu Gly Ala Al - #a Asp Asp Pro Trp Arg                   355          - #       360          - #       365                      - - CTA AGA TCA ACG TAC CAA TCT CAA AAA TAC CA - #G GCT GAA TTC CCC ATG         1328                                                                       Leu Arg Ser Thr Tyr Gln Ser Gln Lys Tyr Gl - #n Ala Glu Phe Pro Met               370              - #   375              - #   380                          - - GGT CCT GTG ACC TCA GCC CAT GCG GGG ACC TA - #C AGG TGC TAC GGC TCA         1376                                                                       Gly Pro Val Thr Ser Ala His Ala Gly Thr Ty - #r Arg Cys Tyr Gly Ser           385                 3 - #90                 3 - #95                 4 -      #00                                                                              - - CAG AGC TCC AAA CCC TAC CTG CTG ACT CAC CC - #C AGT GAC CCC CTG        GAG     1424                                                                    Gln Ser Ser Lys Pro Tyr Leu Leu Thr His Pr - #o Ser Asp Pro Leu Glu                          405  - #               410  - #               415              - - CTC GTG GTC TCA GGA CCG TCT GGG GGC CCC AG - #C TCC CCG ACA ACA GGC         1472                                                                       Leu Val Val Ser Gly Pro Ser Gly Gly Pro Se - #r Ser Pro Thr Thr Gly                       420      - #           425      - #           430                  - - CCC ACC TCC ACA TCT GGC CCT GAG GAC CAG CC - #C CTC ACC CCC ACC GGG         1520                                                                       Pro Thr Ser Thr Ser Gly Pro Glu Asp Gln Pr - #o Leu Thr Pro Thr Gly                   435          - #       440          - #       445                      - - TCG GAT CCC CAG AGT GGT CTG GGA AGG CAC CT - #G GGG GTT GTG ATC GGC         1568                                                                       Ser Asp Pro Gln Ser Gly Leu Gly Arg His Le - #u Gly Val Val Ile Gly               450              - #   455              - #   460                          - - ATC TTG GTG GCC GTC ATC CTA CTG CTC CTC CT - #C CTC CTC CTC CTC TTC         1616                                                                       Ile Leu Val Ala Val Ile Leu Leu Leu Leu Le - #u Leu Leu Leu Leu Phe           465                 4 - #70                 4 - #75                 4 -      #80                                                                              - - CTC ATC CTC CGA CAT CGA CGT CAG GGC AAA CA - #C TGG ACA TCG ACC        CAG     1664                                                                    Leu Ile Leu Arg His Arg Arg Gln Gly Lys Hi - #s Trp Thr Ser Thr Gln                          485  - #               490  - #               495              - - AGA AAG GCT GAT TTC CAA CAT CCT GCA GGG GC - #T GTG GGG CCA GAG CCC         1712                                                                       Arg Lys Ala Asp Phe Gln His Pro Ala Gly Al - #a Val Gly Pro Glu Pro                       500      - #           505      - #           510                  - - ACA GAC AGA CGC CTG CAG TGG AGG TCC AGC CC - #A GCT GCC GAT GCC CAG         1760                                                                       Thr Asp Arg Arg Leu Gln Trp Arg Ser Ser Pr - #o Ala Ala Asp Ala Gln                   515          - #       520          - #       525                      - - GAA GAA AAC CTC TAT GCT GCC GTG AAG CAC AC - #A CAG CCT GAG GAT GGG         1808                                                                       Glu Glu Asn Leu Tyr Ala Ala Val Lys His Th - #r Gln Pro Glu Asp Gly               530              - #   535              - #   540                          - - GTG GAG ATG GAC ACT CGG CAG AGC CCA CAC GA - #T GAA GAC CCC CAG GCA         1856                                                                       Val Glu Met Asp Thr Arg Gln Ser Pro His As - #p Glu Asp Pro Gln Ala           545                 5 - #50                 5 - #55                 5 -      #60                                                                              - - GTG ACG TAT GCC GAG GTG AAA CAC TCC AGA CC - #T AGG AGA GAA ATG        GCT     1904                                                                    Val Thr Tyr Ala Glu Val Lys His Ser Arg Pr - #o Arg Arg Glu Met Ala                          565  - #               570  - #               575              - - TCT CCT CCT TCC CCA CTG TCT GGG GAA TTC CT - #G GAC ACA AAG GAC AGA         1952                                                                       Ser Pro Pro Ser Pro Leu Ser Gly Glu Phe Le - #u Asp Thr Lys Asp Arg                       580      - #           585      - #           590                  - - CAG GCG GAA GAG GAC AGG CAG ATG GAC ACT GA - #G GCT GCT GCA TCT GAA         2000                                                                       Gln Ala Glu Glu Asp Arg Gln Met Asp Thr Gl - #u Ala Ala Ala Ser Glu                   595          - #       600          - #       605                      - - GCC CCC CAG GAT GTG ACC TAC GCC CAG CTG CA - #C AGC TTG ACC CTT AGA         2048                                                                       Ala Pro Gln Asp Val Thr Tyr Ala Gln Leu Hi - #s Ser Leu Thr Leu Arg               610              - #   615              - #   620                          - - CGG AAG GCA ACT GAG CCT CCT CCA TCC CAG GA - #A GGG CCC TCT CCA GCT         2096                                                                       Arg Lys Ala Thr Glu Pro Pro Pro Ser Gln Gl - #u Gly Pro Ser Pro Ala           625                 6 - #30                 6 - #35                 6 -      #40                                                                              - - GTG CCC AGC ATC TAC GCC ACT CTG GCC ATC CA - #C TAG CCCAGGGGGG              2142                                                                      Val Pro Ser Ile Tyr Ala Thr Leu Ala Ile Hi - #s  *                                            645  - #               650                                     - - GACGCAGACC CCACACTCCA TGGAGTCTGG AATGCATGGG AGCTGCCCCC CC -             #AGTGGACA   2202                                                                 - - CCATTGGACC CCACCCAGCC TGGATCTACC CCAGGAGACT CTGGGAACTT TT -            #AGGGGTCA   2262                                                                 - - CTCAATTCTG CAGTATAAAT AACTAATGTC TCTACAATTT TGAAATAAAG CA -            #ACAGACTT   2322                                                                 - - CTCAATAATC AATGAAGTAG CTGAGAAAAC TAAGTCAGAA AGTGCATTAA AC -            #TGAATCAC   2382                                                                 - - AATGTAAATA TTACACATCA AGCGATGAAA CTGGAAAACT ACAAGCCACG AA -            #TGAATGAA   2442                                                                 - - TTAGGAAAGA AAAAAAGTAG GAAATGAATG ATCTTGGCTT TCCTATAAGA AA -            #TTTAGGGC   2502                                                                 - - AGGGCACGGT GGCTCACGCC TGTAATTCCA GCACTTTGGG AGGCCGAGGC GG -            #GCAGATCA   2562                                                                 - - CGAGTTCAGG AGATCGAGAC CATCTTGGCC AACATGGTGA AACCCTGTCT CT -            #CCTAAAAA   2622                                                                 - - TACAAAAATT AGCTGGATGT GGTGGCAGTG CCTGTAATCC CAGCTATTTG GG -            #AGGCTGAG   2682                                                                 - - GCAGGAGAAT CGCTTGAACC AGGGAGTCAG AGGTTTCAGT GAGCCAAGAT CG -            #CACCACTG   2742                                                                 - - CTCTCCAGCC TGGCGACAGA GGGAGACTCC ATCTCAAATT AAAAAAAA  - #                  2790                                                                        - -  - - (2) INFORMATION FOR SEQ ID NO:22:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH:  651 ami - #no acids                                              (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:22:                              - - Met Thr Pro Ile Leu Thr Val Leu Ile Cys Le - #u Gly Leu Ser Leu Gly        1               5 - #                 10 - #                 15              - - Pro Arg Thr His Val Gln Ala Gly His Leu Pr - #o Lys Pro Thr Leu Trp                   20     - #             25     - #             30                  - - Ala Glu Pro Gly Ser Val Ile Thr Gln Gly Se - #r Pro Val Thr Leu Arg               35         - #         40         - #         45                      - - Cys Gln Gly Gly Gln Glu Thr Gln Glu Tyr Ar - #g Leu Tyr Arg Glu Lys           50             - #     55             - #     60                          - - Lys Thr Ala Pro Trp Ile Thr Arg Ile Pro Gl - #n Glu Leu Val Lys Lys       65                 - # 70                 - # 75                 - # 80       - - Gly Gln Phe Pro Ile Pro Ser Ile Thr Trp Gl - #u His Ala Gly Arg Tyr                       85 - #                 90 - #                 95              - - Arg Cys Tyr Tyr Gly Ser Asp Thr Ala Gly Ar - #g Ser Glu Ser Ser Asp                  100      - #           105      - #           110                  - - Pro Leu Glu Leu Val Val Thr Gly Ala Tyr Il - #e Lys Pro Thr Leu Ser              115          - #       120          - #       125                      - - Ala Gln Pro Ser Pro Val Val Asn Ser Gly Gl - #y Asn Val Thr Leu Gln          130              - #   135              - #   140                          - - Cys Asp Ser Gln Val Ala Phe Asp Gly Phe Il - #e Leu Cys Lys Glu Gly      145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - Glu Asp Glu His Pro Gln Cys Leu Asn Ser Gl - #n Pro His Ala Arg        Gly                                                                                             165  - #               170  - #               175             - - Ser Ser Arg Ala Ile Phe Ser Val Gly Pro Va - #l Ser Pro Ser Arg Arg                  180      - #           185      - #           190                  - - Trp Trp Tyr Arg Cys Tyr Ala Tyr Asp Ser As - #n Ser Pro Tyr Glu Trp              195          - #       200          - #       205                      - - Ser Leu Pro Ser Asp Leu Leu Glu Leu Leu Va - #l Leu Gly Val Ser Lys          210              - #   215              - #   220                          - - Lys Pro Ser Leu Ser Val Gln Pro Gly Pro Il - #e Val Ala Pro Glu Glu      225                 2 - #30                 2 - #35                 2 -      #40                                                                              - - Thr Leu Thr Leu Gln Cys Gly Ser Asp Ala Gl - #y Tyr Asn Arg Phe        Val                                                                                             245  - #               250  - #               255             - - Leu Tyr Lys Asp Gly Glu Arg Asp Phe Leu Gl - #n Leu Ala Gly Ala Gln                  260      - #           265      - #           270                  - - Pro Gln Ala Gly Leu Ser Gln Ala Asn Phe Th - #r Leu Gly Pro Val Ser              275          - #       280          - #       285                      - - Arg Ser Tyr Gly Gly Gln Tyr Arg Cys Tyr Gl - #y Ala His Asn Leu Ser          290              - #   295              - #   300                          - - Ser Glu Trp Ser Ala Pro Ser Asp Pro Leu As - #p Ile Leu Ile Ala Gly      305                 3 - #10                 3 - #15                 3 -      #20                                                                              - - Gln Phe Tyr Asp Arg Val Ser Leu Ser Val Gl - #n Pro Gly Pro Thr        Val                                                                                             325  - #               330  - #               335             - - Ala Ser Gly Glu Asn Val Thr Leu Leu Cys Gl - #n Ser Gln Gly Trp Met                  340      - #           345      - #           350                  - - Gln Thr Phe Leu Leu Thr Lys Glu Gly Ala Al - #a Asp Asp Pro Trp Arg              355          - #       360          - #       365                      - - Leu Arg Ser Thr Tyr Gln Ser Gln Lys Tyr Gl - #n Ala Glu Phe Pro Met          370              - #   375              - #   380                          - - Gly Pro Val Thr Ser Ala His Ala Gly Thr Ty - #r Arg Cys Tyr Gly Ser      385                 3 - #90                 3 - #95                 4 -      #00                                                                              - - Gln Ser Ser Lys Pro Tyr Leu Leu Thr His Pr - #o Ser Asp Pro Leu        Glu                                                                                             405  - #               410  - #               415             - - Leu Val Val Ser Gly Pro Ser Gly Gly Pro Se - #r Ser Pro Thr Thr Gly                  420      - #           425      - #           430                  - - Pro Thr Ser Thr Ser Gly Pro Glu Asp Gln Pr - #o Leu Thr Pro Thr Gly              435          - #       440          - #       445                      - - Ser Asp Pro Gln Ser Gly Leu Gly Arg His Le - #u Gly Val Val Ile Gly          450              - #   455              - #   460                          - - Ile Leu Val Ala Val Ile Leu Leu Leu Leu Le - #u Leu Leu Leu Leu Phe      465                 4 - #70                 4 - #75                 4 -      #80                                                                              - - Leu Ile Leu Arg His Arg Arg Gln Gly Lys Hi - #s Trp Thr Ser Thr        Gln                                                                                             485  - #               490  - #               495             - - Arg Lys Ala Asp Phe Gln His Pro Ala Gly Al - #a Val Gly Pro Glu Pro                  500      - #           505      - #           510                  - - Thr Asp Arg Arg Leu Gln Trp Arg Ser Ser Pr - #o Ala Ala Asp Ala Gln              515          - #       520          - #       525                      - - Glu Glu Asn Leu Tyr Ala Ala Val Lys His Th - #r Gln Pro Glu Asp Gly          530              - #   535              - #   540                          - - Val Glu Met Asp Thr Arg Gln Ser Pro His As - #p Glu Asp Pro Gln Ala      545                 5 - #50                 5 - #55                 5 -      #60                                                                              - - Val Thr Tyr Ala Glu Val Lys His Ser Arg Pr - #o Arg Arg Glu Met        Ala                                                                                             565  - #               570  - #               575             - - Ser Pro Pro Ser Pro Leu Ser Gly Glu Phe Le - #u Asp Thr Lys Asp Arg                  580      - #           585      - #           590                  - - Gln Ala Glu Glu Asp Arg Gln Met Asp Thr Gl - #u Ala Ala Ala Ser Glu              595          - #       600          - #       605                      - - Ala Pro Gln Asp Val Thr Tyr Ala Gln Leu Hi - #s Ser Leu Thr Leu Arg          610              - #   615              - #   620                          - - Arg Lys Ala Thr Glu Pro Pro Pro Ser Gln Gl - #u Gly Pro Ser Pro Ala      625                 6 - #30                 6 - #35                 6 -      #40                                                                              - - Val Pro Ser Ile Tyr Ala Thr Leu Ala Ile Hi - #s                                          645  - #               650                                  __________________________________________________________________________

What is claimed is:
 1. An isolated polypeptide comprising at least 12contiguous amino acid residues within a mature DLAIR amino acid sequenceselected from the group consisting of residues +1 to 266 of SEQ ID NO: 6and residues +1 to 114 of SEQ ID NO:
 10. 2. A polypeptide according toclaim 1 comprising at least 14 contiguous amino acids of said matureDLAIR amino acid sequence.
 3. A polypeptide according to claim 1comprising at least 16 contiguous amino acids of said mature DLAIR aminoacid sequence.
 4. A polypeptide according to claim 1 comprising at least18 contiguous amino acids of said mature DLAIR amino acid sequence.
 5. Apolypeptide according to claim 1 comprising at least 20 contiguous aminoacids of said mature DLAIR amino acid sequence.
 6. A polypeptideaccording to claim 1 comprising at least 22 contiguous amino acids ofsaid mature DLAIR amino acid sequence, wherein the polypeptidespecifically binds an antibody raised against a polypeptide consistingof said mature DLAIR amino acid sequence.
 7. A polypeptide according toclaim 1 comprising at least 30 contiguous amino acids of said matureDLAIR amino acid sequence.
 8. An isolated polypeptide comprising theamino acid sequence shown as residues +1 to 266 of SEQ ID NO:
 6. 9. Anisolated polypeptide comprising the amino acid sequence shown asresidues +1 to 114 of SEQ ID NO:
 10. 10. A polypeptide according toclaim 1 further comprising a heterologous amino acid sequence.
 11. Apolypeptide according to claim 8 further comprising a heterologous aminoacid sequence.
 12. A polypeptide according to claim 9 further comprisinga heterologous amino acid sequence.
 13. A polypeptide according to claim10, wherein the heterologous amino acid sequence is a detection orpurification tag sequence.
 14. A polypeptide according to claim 11,wherein the heterologous amino acid sequence is a detection orpurification tag sequence.
 15. A polypeptide according to claim 12,wherein the heterologous amino acid sequence is a detection orpurification tag sequence.
 16. A composition comprising a polypeptideaccording to claim 1 and a solvent or carrier.
 17. A compositioncomprising a polypeptide according to claim 8 and a solvent or carrier.18. A composition comprising a polypeptide according to claim 9 and asolvent or carrier.
 19. An isolated nucleic acid molecule comprising asequence encoding at least 12 contiguous amino acid residues within themature DLAIR amino acid sequence shown as residues +1 to 266 of SEQ IDNO:
 6. 20. A nucleic acid molecule according to claim 19 comprising asequence encoding at least 14 contiguous amino acids of said matureDLAIR amino acid sequence.
 21. A nucleic acid molecule according toclaim 19 comprising a sequence encoding at least 16 contiguous aminoacids of said mature DLAIR amino acid sequence.
 22. A nucleic acidmolecule according to claim 19 comprising a sequence encoding at least18 contiguous amino acids of said mature DLAIR amino acid sequence. 23.A nucleic acid molecule according to claim 19 comprising a sequenceencoding at least 20 contiguous amino acids of said mature DLAIR aminoacid sequence.
 24. A nucleic acid molecule according to claim 19comprising a sequence encoding at least 22 contiguous amino acids ofsaid mature DLAIR amino acid sequence, wherein the nucleic acid moleculeencodes a polypeptide that specifically binds an antibody raised againsta polypeptide consisting of said mature DLAIR amino acid sequence.
 25. Anucleic acid molecule according to claim 19 comprising a sequenceencoding at least 30 contiguous amino acids of said mature DLAIR aminoacid sequence.
 26. A nucleic acid molecule according to claim 19,wherein the sequence encoding said at least 12 amino acid residues is acontiguous fragment of the nucleotide sequence shown in SEQ ID NO: 5 orof a sequence which differs therefrom only by the replacement of Tresidues by U residues.
 27. A nucleic acid molecule according to claim26 comprising at least 50 contiguous nucleotides of SEQ ID NO: 5 or of asequence which differs therefrom only by the replacement of T residuesby U residues.
 28. A nucleic acid molecule according to claim 27,wherein the nucleic acid molecule encodes a fusion protein comprising atleast 24 contiguous amino acid residues of SEQ ID NO: 6 and aheterologous amino acid sequence.
 29. A nucleic acid molecule accordingto claim 26 comprising at least 75 contiguous nucleotides of SEQ ID NO:5 or of a sequence which differs therefrom only by the replacement of Tresidues by U residues.
 30. A nucleic acid molecule according to claim26 comprising at least 100 contiguous nucleotides of SEQ ID NO: 5 or ofa sequence which differs therefrom only by the replacement of T residuesby U residues.
 31. An isolated nucleic acid molecule encoding apolypeptide comprising the amino acid sequence shown as residues +1 to266 of SEQ ID NO:
 6. 32. A nucleic acid molecule according to claim 31,comprising the nucleotide sequence shown as residues 218 to 1015 of SEQID NO: 5 or a sequence which differs therefrom only by the replacementof T residues by U residues.
 33. An isolated nucleic acid moleculeencoding a polypeptide comprising the amino acid sequence shown asresidues +1 to 114 of SEQ ID NO:
 10. 34. A nucleic acid moleculeaccording to claim 33, comprising the nucleotide sequence shown asresidues 87 to 428 of SEQ ID NO: 9 or a sequence which differs therefromonly by the replacement of T residues by U residues.
 35. A nucleic acidmolecule according to claim 19 further comprising a sequence encoding aheterologous amino acid sequence.
 36. A nucleic acid molecule accordingto claim 31 further comprising a sequence encoding a heterologous aminoacid sequence.
 37. A nucleic acid molecule according to claim 33 furthercomprising a sequence encoding a heterologous amino acid sequence.
 38. Anucleic acid molecule according to claim 35, wherein the heterologousamino acid sequence is a detection or purification tag sequence.
 39. Anucleic acid molecule according to claim 36, wherein the heterologousamino acid sequence is a detection or purification tag sequence.
 40. Anucleic acid molecule according to claim 32, wherein the heterologousamino acid sequence is a detection or purification tag sequence.
 41. Amethod of making a nucleic acid duplex, comprising contacting a firstnucleic acid molecule according to claim 19 with a second nucleic acidmolecule under hybridization conditions of at least 45° C. and less than500 mM salt, wherein said first and second nucleic acid molecules form aduplex under said conditions.
 42. A method of making a nucleic acidmolecule according to claim 19, comprising:providing DNA or RNAcomprising the sequence of the molecule; and amplifying the sequence bythe polymerase chain reaction.
 43. A method of making a nucleic acidmolecule according to claim 31, comprising:providing DNA or RNAcomprising the sequence of the molecule; and amplifying the sequence bythe polymerase chain reaction.
 44. A method of making a nucleic acidmolecule according to claim 33, comprising:providing DNA or RNAcomprising the sequence of the molecule; and amplifying the sequence bythe polymerase chain reaction.
 45. An expression vector having a codingsequence that comprises:(a) a sequence encoding at least 12 contiguousamino acid residues within residues +1 to 266 of SEQ ID NO: 6; or (b) asequence encoding at least 12 contiguous amino acid residues withinresidues +1 to 114 of SEQ ID NO:
 10. 46. An expression vector accordingto claim 45 comprising:(a) a sequence encoding at least 30 contiguousamino acid residues within residues +1 to 266 of SEQ ID NO: 6; or (b) asequence encoding at least 30 contiguous amino acid residues withinresidues +1 to 114 of SEQ ID NO:
 10. 47. An expression vector accordingto claim 45 that comprises:(a) at least 50 contiguous nucleotideresidues within residues 218 to 1015 of SEQ ID NO: 5; (b) at least 50contiguous nucleotide residues within residues 87 to 428 of SEQ ID NO:9; or (c) a sequence which differs from the 50 contiguous residues of(a) or (b) only by the replacement of T residues by U residues.
 48. Anexpression vector according to claim 45 comprising:(a) at least 75contiguous nucleotide residues within residues 218 to 1015 of SEQ ID NO:5; (b) at least 75 contiguous nucleotide residues within residues 87 to428 of SEQ ID NO: 9; or (c) a sequence which differs from the 75contiguous residues of (a) or (b) only by the replacement of T residuesby U residues.
 49. An expression vector according to claim 45comprising:(a) at least 100 contiguous nucleotide residues withinresidues 218 to 1015 of SEQ ID NO: 5; (b) at least 100 contiguousnucleotide residues within residues 87 to 428 of SEQ ID NO: 9; or (c) asequence which differs from the 100 contiguous residues of (a) or (b)only by the replacement of T residues by U residues.
 50. An expressionvector according to claim 45 comprising a sequence that encodes apolypeptide comprising the amino acid sequence shown as residues +1 to266 of SEQ ID NO:
 6. 51. An expression vector according to claim 45comprising a sequence that encodes a polypeptide comprising the aminoacid sequence shown as residues +1 to 114 of SEQ ID NO:
 10. 52. A cellcomprising an expression vector according to claim
 45. 53. A cellaccording to claim 52, wherein the cell is a prokaryotic cell.
 54. Acell according to claim 52, wherein the cell is a eukaryotic cell.
 55. Acell according to claim 54, wherein the cell is a yeast cell or aninsect cell.
 56. A cell according to claim 54, wherein the cell is arodent cell or a human cell.
 57. A method of making a polypeptide,comprising culturing a cell according to claim 52 under conditionssuitable to effect the expression of the polypeptide encoded by thecoding sequence in the expression vector.
 58. A method according toclaim 57, wherein the polypeptide comprises the amino acid sequenceshown as residues +1 to 266 of SEQ ID NO:
 6. 59. A method according toclaim 57, wherein the polypeptide comprises the amino acid sequenceshown as residues +1 to 114 of SEQ ID NO:
 10. 60. A method according toclaim 58, wherein the polypeptide further comprises a heterologous aminoacid sequence.
 61. A method according to claim 59, wherein thepolypeptide further comprises a heterologous amino acid sequence.